Updated on 2026/03/05

写真a

 
MIE Masayasu
 
Organization
School of Life Science and Technology Associate Professor
Title
Associate Professor
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Degree

  • Doctor of Engineering ( Tokyo Institute of Technology )

Research Areas

  • Life Science / Biomedical engineering

  • Life Science / Biomaterials

Education

  • Tokyo Institute of Technology   Bioscience and Biotechnology

    - 2000

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    Country： Japan

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  • Tokyo Institute of Technology   School of Bioscience and Biotechnology

    - 1995

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    Country： Japan

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Research History

  • Institute of Science Tokyo   Associate Professor

    2025.10

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  • Tokyo Institute of Technology   Associate Professor

    2016.4 - 2024.9

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  • Tokyo Institute of Technology   Associate Professor

    2013.12 - 2016.3

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  • Tokyo Institute of Technology   Assistant Professor

    2007.4 - 2013.11

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  • Tokyo Institute of Technology

    2001.4 - 2007.3

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  • 日本学術振興会 特別研究員(PD)

    2000.4 - 2001.3

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  • 日本学術振興会 特別研究員(DC)

    1999.1 - 2000.3

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Professional Memberships

Papers

  • Magnetic Cell Separation Based on Protein Nanoparticles Mediating the Interaction between Magnetic Particles and Target Cells. Reviewed International journal

    Kei Nishida, Gaoyang Wang, Eiry Kobatake, Masayasu Mie

    ACS applied bio materials   8 ( 2 )   1126 - 1137   2025.2

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    Authorship：Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

    Isolation of specific cells from biological samples is an important aspect of various biological research and diagnostic applications. Magnetic separation using magnetic particles (MPs) allows for easy and specific isolation of the target cells. However, depending on the target cell antigen, biological ligands, such as antibodies, must be modified or altered on MPs. Additionally, further biological evaluation of isolated cells requires the removal of MPs from cells by the enzymatic degradation of the biological ligands. In this study, we designed a magnetic cell separation system in which temperature-responsive protein nanoparticles mediated the interaction between target cells and MPs, achieving the easy changeability of biological ligands, removal of MPs by cooling, and effective cell isolation. The protein nanoparticles were thermally responsively formed from fusion proteins constituted of elastin-like polypeptide (ELP), poly(aspartic acid) [poly(d)], and proteins-of-interest such as NanoLuc luciferase (Nluc) fused with replication initiation protein (Rep) (ELP-poly(d)-Nluc-Rep) or biotin acceptor peptide (BAP) (ELP-poly(d)-Nluc-BAP). Rep exhibited enzymatic conjugation activity with an optional DNA aptamer to protein nanoparticles. The transmembrane glycoprotein mucin 1 (MUC1)-binding DNA aptamer was conjugated to Rep as a model aptamer. Bioluminescence signals emitted from the Nluc domains were used to analyze the binding abilities. BAP contributed to binding to streptavidin-modified MPs via a biotin-streptavidin interaction. The MUC1-conjugated protein nanoparticles bound to MUC1-positive human breast cancer MCF-7 cells via MUC1 aptamers and streptavidin-conjugated MPs via BAP, leading to magnetic cell separation. The ratio of isolated MCF-7 cells via magnetic separation was 71.3% for the MCF-7 suspension at 1000 cells/1 mL. The MPs bound on recovered MCF-7 cells were removed by cooling at 4 °C to induce the dissociation of protein nanoparticles. Magnetic cell separation systems that use protein nanoparticles are a promising technology for biological research and diagnostic applications.

    DOI： 10.1021/acsabm.4c01450

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  • Cholesterol- and ssDNA-binding fusion protein-mediated DNA tethering on the plasma membrane. Reviewed International journal

    Kei Nishida, Minon Ishizuka, Eiry Kobatake, Masayasu Mie

    Biomaterials science   13 ( 1 )   299 - 309   2024.12

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    Authorship：Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

    DNA modification of the plasma membrane is an excellent approach for controlling membrane-protein interactions, modulating cell-cell/cell-biomolecule interactions, and extending the biosensing field. The hydrophobic insertion of DNA conjugated with hydrophobic anchoring molecules is utilized for tethering DNA on the cell membrane. In this study, we developed an alternative approach to tether DNA on the plasma membrane based on ssDNA- and cholesterol-binding proteins. We designed a fusion protein (Rep-ALOD4) composed of domain 4 of anthrolysin O (ALOD4), which binds to cholesterol in the plasma membrane, and a replication initiator protein derived from porcine circovirus type 2 (Rep), which forms covalent bonds with single-stranded DNA (ssDNA) with a Rep recognition sequence. Rep-ALOD4 conjugates ssDNA to Rep and binds to the plasma membrane via cholesterol, thus tethering ssDNA to the cells. Quartz crystal microbalance measurements showed that membrane cholesterol binding of Rep-ALOD4 to the lipid bilayer containing cholesterol was accelerated above 20% (w/w) cholesterol in the lipid bilayer. Rep-ALOD4 was conjugated to fluorescein-labeled ssDNA (S-FITC-Rep-ALOD4) and used to treat human cervical tumor HeLa cells. The green signal assigned to S-FITC-Rep-ALOD4 was detected along HeLa cells, whereas diminished by cholesterol removal with methyl β-cyclodextrins. Moreover, ssDNA-conjugated Rep-ALOD4 tethered ssDNA-conjugated functional proteins on the HeLa cell plasma membrane via complementary base pairing. Collectively, Rep-ALOD4 has the potential as an ssDNA-tethering material via plasma membrane cholesterol to extend cell surface engineering.

    DOI： 10.1039/d4bm01127a

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  • Functional decoration of elastin-like polypeptides-based nanoparticles with a modular assembly via isopeptide bond formation Reviewed International journal

    Jun Yamaguchi, Kei Nishida, Eiry Kobatake, Masayasu Mie

    Biotechnology Letters   47 ( 1 )   2024.11

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    Authorship：Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：Springer Science and Business Media LLC  

    DOI： 10.1007/s10529-024-03549-1

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    Other Link： https://link.springer.com/article/10.1007/s10529-024-03549-1/fulltext.html

  • Sensitive Detection of Tumor Cells Using Protein Nanoparticles with Multiple Displays of DNA Aptamers and Bioluminescent Reporters Reviewed International journal

    Kei Nishida, Gaoyang Wang, Eiry Kobatake, Masayasu Mie

    ACS Biomaterials Science & Engineering   2023.9

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    Authorship：Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

    DOI： 10.1021/acsbiomaterials.3c00712

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  • Development of an enhanced immunoassay based on protein nanoparticles displaying an IgG-binding domain and luciferase. Reviewed International journal

    Gaoyang Wang, Yasumasa Mashimo, Eiry Kobatake, Masayasu Mie

    Analytical and bioanalytical chemistry   414 ( 6 )   2079 - 2088   2022.3

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    Authorship：Corresponding author   Language：English   Publishing type：Research paper (scientific journal)  

    Detection of small amounts of target molecules with high sensitivity is important for the diagnosis of many diseases, including cancers, and is particularly important to detect early stages of disease. Here, we report the development of a temperature-responsive fusion protein (ELP-DCN) comprised of an elastin-like polypeptide (ELP), poly-aspartic acid (D), antibody-binding domain C (C), and NanoLuc luciferase (N). ELP-DCN proteins form nanoparticles above a certain threshold temperature that display an antibody-binding domain and NanoLuc luciferase on their surface. ELP-DCN nanoparticles can be applied for enhancement of immunoassay systems because they provide more antibody-binding sites and an increased number of luciferase molecules, resulting in an increase in assay signal. Here, we report the detection of human serum albumin (HSA) as a model protein using anti-HSA and ELP-DCN proteins. Upon formation of ELP-DCN nanoparticles, the detection limit improved tenfold compared to the monomeric form of ELP-DCN.

    DOI： 10.1007/s00216-021-03842-2

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  • Antibody response to the first dose of AZD1222 vaccine in COVID-19 convalescent and uninfected individuals in Bangladesh. Reviewed International coauthorship International journal

    Raeed Jamiruddin, Ahsanul Haq, Mohib Ullah Khondoker, Tamanna Ali, Firoz Ahmed Md, Shahad Saif Khandker, Irfan Jawad, Rubel Hossain, Sohel Ahmed, Sabita Rezwana Rahman, Mamun Mustafi, Taku Kaitsuka, Masayasu Mie, Kazuhito Tomizawa, Eiry Kobatake, Mainul Haque, Nihad Adnan

    Expert review of vaccines   2021.9

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    Language：English   Publishing type：Research paper (scientific journal)  

    BACKGROUND: Vaccination with the Oxford-AstraZeneca COVID-19 vaccine (AZD1222) initially started in the UK and quickly implemented around the Globe, including Bangladesh. Up to date, more than nine million doses administrated to the Bangladeshi public. METHOD: Herein, we studied the antibody response to the first dose of AZD1222 in 86 Bangladeshi individuals using in-house ELISA kits. Study subjects were categorized into two groups, convalescent and uninfected, based on prior infection history and SARS-CoV-2 nucleocapsid-IgG profiles. RESULTS: All the convalescent individuals presented elevated spike-1-IgG compared to 90% of uninfected ones after the first dose. Day >28 post-vaccination, the convalescent group showed six times higher antibody titer than the uninfected ones. The most elevated antibody titers for the former and later group were found at Day 14 and Days >28 post-vaccination, respectively. The spike-1-IgA titer showed a similar pattern as spike-1-IgG, although in a low-titer. In contrast, the IgM titer did not show any significant change in either group. CONCLUSION: High antibody titer in the convalescent group, signify the importance of the first dose among the uninfected group. This study advocates the integration of antibody tests in vaccination programs in the healthcare system for maximizing benefit.

    DOI： 10.1080/14760584.2021.1977630

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  • Detection of SARS-CoV-2 by antigen ELISA test is highly swayed by viral load and sample storage condition. Reviewed International coauthorship International journal

    Nihad Adnan, Shahad Saif Khandker, Ahsanul Haq, Mousumi Akter Chaity, Abdul Khalek, Anawarul Quader Nazim, Taku Kaitsuka, Kazuhito Tomizawa, Masayasu Mie, Eiry Kobatake, Sohel Ahmed, Nor Azlina A Rahman, Mohib Ullah Khondoker, Mainul Haque, Mohd Raeed Jamiruddin

    Expert review of anti-infective therapy   2021.9

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    Language：English   Publishing type：Research paper (scientific journal)  

    BACKGROUND: Rapid increase in COVID-19 suspected cases has rendered disease diagnosis challenging, mainly depending upon RT-qPCR. Reliable, rapid, and cost-effective diagnostic assays that complement RT-qPCR should be introduced after thoroughly evaluating their performance upon various disease phases, viral load, and storage conditions. OBJECTIVE: We investigated the correlation of cycle threshold (Ct) value, which implies the viral load and infection phase, and the storage condition of the clinical specimen with the diagnosis of SARS-CoV-2 through our newly developed in-house rapid enzyme-linked immunosorbent assay (ELISA) system. METHOD: Naso-oropharyngeal samples of 339 COVID-19 suspected cases were collected and evaluated through RT-qPCR that were stored up to 30 days in different conditions (i.e., -80°C, -20°C and initially at 4°C followed by -80°C). The clinical specimens were evaluated with our in-house ELISA system after finalizing the assay method through checkerboard assay and minimizing the signal/noise ratio. RESULT: The ELISA system showed the highest sensitivity (92.9%) for samples with Ct ≤30 and preserving at -80°C temperature. The sensitivity reduced proportionally with increasing Ct value and preserving temperature. However, the specificity ranged between 98.3-100%. CONCLUSION: The results indicate the necessity of early infection phase diagnosis and lower temperature preservation of samples to perform rapid antigen ELISA tests.

    DOI： 10.1080/14787210.2021.1976144

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  • Longitudinal Antibody Dynamics Against Structural Proteins of SARS-CoV-2 in Three COVID-19 Patients Shows Concurrent Development of IgA, IgM, and IgG. Reviewed International coauthorship International journal

    Mohd Raeed Jamiruddin, Md Ahsanul Haq, Kazuhito Tomizawa, Eiry Kobatake, Masayasu Mie, Sohel Ahmed, Shahad Saif Khandker, Tamanna Ali, Nowshin Jahan, Mumtarin Jannat Oishee, Mohib Ullah Khondoker, Bijon Kumar Sil, Mainul Haque, Nihad Adnan

    Journal of inflammation research   14   2497 - 2506   2021

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    Language：English   Publishing type：Research paper (scientific journal)  

    Background: Dynamics and persistence of neutralizing and non-neutralizing antibodies can give us the knowledge required for serodiagnosis, disease management, and successful vaccine design and development. The disappearance of antibodies, absence of humoral immunity activation, and sporadic reinfection cases emphasize the importance of longitudinal antibody dynamics against variable structural antigens. Methods: In this study, twenty-five healthy subjects working in a SARS-COV-2 serodiagnostic assay development project were enrolled, and their sign and symptoms were followed up to six months. Three subjects showed COVID-19-like symptoms, and three subjects' antibody dynamics were followed over 120 days by analyzing 516 samples. We have developed 12 different types of in-house ELISAs to observe the kinetics of IgG, IgM, and IgA against four SARS-CoV-2 proteins, namely nucleocapsid, RBD, S1, and whole spike (S1+S2). For the development of these assays, 30-104 pre-pandemic samples were taken as negative controls and 83 RT-qPCR positive samples as positive ones. Results: All three subjects presented COVID-19-like symptoms twice, with mild symptoms in the first episode were severe in the second, and RT-qPCR confirmed the latter. The initial episode did not culminate with any significant antibody development, while a multifold increase in IgG antibodies characterized the second episode. Interestingly, IgG antibody development concurrent with IgM and IgA and persisted, whereas the latter two weans off rather quickly if appeared. Conclusion: Antibody kinetics observed in this study can provide a pathway to the successful development of sero-diagnostics and epidemiologists to predict the fate of vaccination currently in place.

    DOI： 10.2147/JIR.S313188

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  • AuNP Coupled Rapid Flow-Through Dot-Blot Immuno-Assay for Enhanced Detection of SARS-CoV-2 Specific Nucleocapsid and Receptor Binding Domain IgG. Reviewed International coauthorship International journal

    Bijon Kumar Sil, Mohd Raeed Jamiruddin, Md Ahsanul Haq, Mohib Ullah Khondoker, Nowshin Jahan, Shahad Saif Khandker, Tamanna Ali, Mumtarin Jannat Oishee, Taku Kaitsuka, Masayasu Mie, Kazuhito Tomizawa, Eiry Kobatake, Mainul Haque, Nihad Adnan

    International journal of nanomedicine   16   4739 - 4753   2021

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    Language：English   Publishing type：Research paper (scientific journal)  

    Background: Serological tests detecting severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) are widely used in seroprevalence studies and evaluating the efficacy of the vaccination program. Some of the widely used serological testing techniques are enzyme-linked immune-sorbent assay (ELISA), chemiluminescence immunoassay (CLIA), and lateral flow immunoassay (LFIA). However, these tests are plagued with low sensitivity or specificity, time-consuming, labor-intensive, and expensive. We developed a serological test implementing flow-through dot-blot assay (FT-DBA) for SARS-CoV-2 specific IgG detection, which provides enhanced sensitivity and specificity while being quick to perform and easy to use. Methods: SARS-CoV-2 antigens were immobilized on nitrocellulose membrane to capture human IgG, which was then detected with anti-human IgG conjugated gold nanoparticle (hIgG-AuNP). A total of 181 samples were analyzed in-house. Within which 35 were further evaluated in US FDA-approved CLIA Elecsys SARS-CoV-2 assay. The positive panel consisted of RT-qPCR positive samples from patients with both <14 days and >14 days from the onset of clinical symptoms. The negative panel contained samples collected from the pre-pandemic era dengue patients and healthy donors during the pandemic. Moreover, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of FT-DBA were evaluated against RT-qPCR positive sera. However, the overall efficacies were assessed with sera that seroconverted against either nucleocapsid (NCP) or receptor-binding domain (RBD). Results: In-house ELISA selected a total of 81 true seropositive and 100 seronegative samples. The sensitivity of samples with <14 days using FT-DBA was 94.7%, increasing to 100% for samples >14 days. The overall detection sensitivity and specificity were 98.8% and 98%, respectively, whereas the overall PPV and NPV were 99.6% and 99%. Moreover, comparative analysis between in-house ELISA assays and FT-DBA revealed clinical agreement of Cohen's Kappa value of 0.944. The FT-DBA showed sensitivity and specificity of 100% when compared with commercial CLIA kits. Conclusion: The assay can confirm past SARS-CoV-2 infection with high accuracy within 2 minutes compared to commercial CLIA or in-house ELISA. It can help track SARS-CoV-2 disease progression, population screening, and vaccination response. The ease of use of the assay without requiring any instruments while being semi-quantitative provides the avenue of its implementation in remote areas around the globe, where conventional serodiagnosis is not feasible.

    DOI： 10.2147/IJN.S313140

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  • Development and performance evaluation of a rapid in-house ELISA for retrospective serosurveillance of SARS-CoV-2. International journal

    Bijon Kumar Sil, Nowshin Jahan, Md Ahsanul Haq, Mumtarin Jannat Oishee, Tamanna Ali, Shahad Saif Khandker, Eiry Kobatake, Masayasu Mie, Mohib Ullah Khondoker, Mohd Raeed Jamiruddin, Nihad Adnan

    PloS one   16 ( 2 )   e0246346   2021

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    Language：English   Publishing type：Research paper (scientific journal)  

    BACKGROUND: In the ongoing pandemic situation of COVID-19, serological tests can complement the molecular diagnostic methods, and can be one of the important tools of sero-surveillance and vaccine evaluation. AIM: To develop and evaluate a rapid SARS-CoV-2 specific ELISA for detection of anti-SARS-CoV2 IgG from patients' biological samples. METHODS: In order to develop this ELISA, three panels of samples (n = 184) have been used: panel 1 (n = 19) and panel 2 (n = 60) were collected from RT-PCR positive patients within 14 and after 14 days of onset of clinical symptoms, respectively; whereas panel 3 consisted of negative samples (n = 105) collected either from healthy donors or pre-pandemic dengue patients. As a capturing agent full-length SARS-CoV2 specific recombinant nucleocapsid was immobilized. Commercial SARS-CoV2 IgG kit based on chemiluminescent assay was used for the selection of samples and optimization of the assay. The threshold cut-off point, inter-assay and intra-assay variations were determined. RESULTS: The incubation/reaction time was set at a total of 30 minutes with the sensitivity of 84% (95% confidence interval, CI, 60.4%, 96.6%) and 98% (95% CI, 91.1%, 100.0%), for panel 1 and 2, respectively; with overall 94.9% sensitivity (95% CI 87.5%, 98.6%). Moreover, the clinical specificity was 97.1% (95% CI, 91.9%, 99.4%) with no cross reaction with dengue samples. The overall positive and negative predictive values are 96.2% (95% CI 89.2%, 99.2%) and 96.2% (95% CI, 90.6% 99.0%), respectively. In-house ELISA demonstrated 100% positive and negative percent agreement with Elecsys Anti-SARS-CoV-2, with Cohen's kappa value of 1.00 (very strong agreement), while comparing 13 positive and 17 negative confirmed cases. CONCLUSION: The assay is rapid and can be applied as one of the early and retrospective sero-monitoring tools in all over the affected areas.

    DOI： 10.1371/journal.pone.0246346

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  • Construction of an Enzymatically-Conjugated DNA Aptamer–Protein Hybrid Molecule for Use as a BRET-Based Biosensor Reviewed International journal

    Masayasu Mie, Rena Hirashima, Yasumasa Mashimo, Eiry Kobatake

    Applied Sciences   10 ( 21 )   7646 - 7646   2020.10

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    Authorship：Corresponding author   Publishing type：Research paper (scientific journal)   Publisher：{MDPI} {AG}  

    DOI： 10.3390/app10217646

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  • Construction of multifunctional fusion proteins with a laminin-derived short peptide to promote neural differentiation of mouse induced pluripotent stem cells Reviewed International journal

    Afroza Sharmin, Nihad Adnan, Amranul Haque, Yasumasa Mashimo, Masayasu Mie, Eiry Kobatake

    Journal of Biomedical Materials Research - Part B Applied Biomaterials   108 ( 6 )   2691 - 2698   2020.8

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    Language：English   Publishing type：Research paper (scientific journal)   Publisher：John Wiley and Sons Inc.  

    DOI： 10.1002/jbm.b.34600

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  • Construction of DNA-displaying nanoparticles by enzymatic conjugation of DNA and elastin-like polypeptides using a replication initiation protein Reviewed International journal

    Wei Guo, Yasumasa Mashimo, Eiry Kobatake, Masayasu Mie

    Nanotechnology   2020.4

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    Authorship：Corresponding author   Publishing type：Research paper (scientific journal)   Publisher：{IOP} Publishing  

    DOI： 10.1088/1361-6528/ab8042

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  • Temperature-Responsive Multifunctional Protein Hydrogels with Elastin-like Polypeptides for 3-D Angiogenesis Reviewed International journal

    Yoshinori Mizuguchi, Yasumasa Mashimo, Masayasu Mie, Eiry Kobatake

    Biomacromolecules   21 ( 3 )   1126 - 1135   2020.3

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    Language：English   Publishing type：Research paper (scientific journal)   Publisher：American Chemical Society  

    DOI： 10.1021/acs.biomac.9b01496

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  • Direct labeling of protein nanoparticles with fluorescent compounds for immunoassay applications Reviewed International journal

    Tsutomu Sugihara, Masayasu Mie, Eiry Kobatake

    Analytical Sciences   36 ( 3 )   385 - 387   2020

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    Language：English   Publishing type：Research paper (scientific journal)   Publisher：Japan Society for Analytical Chemistry  

    DOI： 10.2116/analsci.19N024

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  • Construction of DNA-NanoLuc luciferase conjugates for DNA aptamer-based sandwich assay using Rep protein Reviewed International journal

    Masayasu Mie, Takahiro Niimi, Yasumasa Mashimo, Eiry Kobatake

    Biotechnology Letters   41 ( 3 )   357 - 362   2019.3

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    Authorship：Lead author,　Corresponding author   Publishing type：Research paper (scientific journal)   Publisher：Springer Science and Business Media {LLC}  

    DOI： 10.1007/s10529-018-02641-7

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  • Development of drug-loaded protein nanoparticles displaying enzymatically-conjugated DNA aptamers for cancer cell targeting Reviewed International coauthorship International journal

    Masayasu Mie, Rie Matsumoto, Yasumasa Mashimo, Anthony E. G. Cass, Eiry Kobatake

    Molecular Biology Reports   46 ( 1 )   261 - 269   2019.2

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    Language：English   Publishing type：Research paper (scientific journal)   Publisher：Springer Netherlands  

    DOI： 10.1007/s11033-018-4467-2

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  • Application of elastin-based nanoparticles displaying antibody binding domains for a homogeneous immunoassay Reviewed International journal

    Tsutomu Sugihara, Masayasu Mie, Eiry Kobatake

    Analytical Biochemistry   544   72 - 79   2018.3

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    Publishing type：Research paper (scientific journal)   Publisher：Elsevier {BV}  

    DOI： 10.1016/j.ab.2017.12.023

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  • A DNA-scaffold platform enhances a multi-enzymatic cycling reaction. Reviewed International journal

    Mashimo, Yasumasa, Mie, Masayasu, Kobatake, Eiry

    Biotechnology letters   2018

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1007/s10529-018-2517-4

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  • Fluorescent and luminescent fusion proteins for analyses of amyloid beta peptide aggregation Reviewed International journal

    Usui, Kenji, Mie, Masayasu, Andou, Takashi, Mihara, Hisakazu, Kobatake, Eiry

    Journal of Peptide Science   23 ( 7-8 )   659 - 665   2017.7

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    DOI： 10.1002/psc.3003

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  • Design of bFGF-tethered self-assembling extracellular matrix proteins via coiled-coil triple-helix formation Reviewed International journal

    Mizuguchi, Yoshinori, Mashimo, Yasumasa, Mie, Masayasu, Kobatake, Eiry

    Biomedical Materials   12 ( 4 )   2017

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1088/1748-605X/aa7616

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  • Construction of a Defined Biomimetic Matrix for Long-Term Maintenance of Mouse Induced Pluripotent Stem Cells

    Adnan, Nihad, Mie, Masayasu, Hague, Amranul, Hossain, Sharif, Mashimo, Yasumasa, Akaike, Toshihiro, Kobatake, Eiry

    Bioconjugate Chemistry   27 ( 7 )   2016

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1021/acs.bioconjchem.6b00141

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  • Design of luciferase-displaying protein nanoparticles for use as highly sensitive immunoassay detection probes

    Ikeda, Yusuke, Mashimo, Yasumasa, Mie, Masayasu, Kobatake, Eiry

    Analyst   141 ( 24 )   2016

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1039/c6an01253a

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  • Construction of a tissue-specific transcription factor-tethered extracellular matrix protein via coiled-coil helix formation

    Siew, SokeLee, Kaneko, Mami, Mie, Masayasu, Kobatake, Eiry

    Journal of Materials Chemistry B   4 ( 14 )   2016

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1039/c5tb01579k

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  • Development of a Split SNAP-CLIP Double Labeling System for Tracking Proteins Following Dissociation from Protein-Protein Complexes in Living Cells.

    Mie, Masayasu, Naoki, Tatsuhiko, Kobatake, Eiry

    Analytical chemistry   88 ( 16 )   2016

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1021/acs.analchem.6b01906

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  • Delivery of bFGF for Tissue Engineering by Tethering to the ECM.

    Suttinont, Chawapun, Mashimo, Yasumasa, Mie, Masayasu, Kobatake, Eiry

    BioMed research international   2015   208089 - 208089   2015

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1155/2015/208089

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  • Growth Factor Tethering to Protein Nanoparticles via Coiled-Coil Formation for Targeted Drug Delivery

    Assal, Yasmine, Mizuguchi, Yoshinori, Mie, Masayasu, Kobatake, Eiry

    Bioconjugate Chemistry   26 ( 8 )   2015

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1021/acs.bioconjchem.5b00266

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  • Tracking a protein following dissociation from a protein-protein complex using a split SNAP-tag system

    Mie, Masayasu, Naoki, Tatsuhiko, Kobatake, Eiry

    Analytical Biochemistry   477   2015

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1016/j.ab.2015.02.019

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  • Assembly of zinc finger motif-fused enzymes on a dsDNA scaffold for catalyzing consecutive reactions with a proximity effect Reviewed

    Funabashi, Hisakage, Yanagi, Satoshi, Suzuki, Shigeya, Mie, Masayasu, Kobatake, Eiry

    Biotechnology Letters   37 ( 1 )   109 - 114   2015

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1007/s10529-014-1644-9

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  • Cellular differentiation assessments by measuring the degree of cellular internalization and membrane adsorption using designed peptides Reviewed

    Usui, Kenji, Kikuchi, Takuya, Kikuchi, Kunio, Mie, Masayasu, Kobatake, Eiry, Mihara, Hisakazu

    Bioorganic & Medicinal Chemistry Letters   24 ( 17 )   4129 - 4131   2014.9

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    Language：English   Publishing type：Research paper (scientific journal)  

    DOI： 10.1016/j.bmcl.2014.07.053

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  • Targeting of EGF-displayed protein nanoparticles with anticancer drugs

    Matsumoto, Rie, Nara, Rieko, Andou, Takashi, Mie, Masayasu, Kobatake, Eiry

    Journal of Biomedical Materials Research Part B-Applied Biomaterials   102 ( 8 )   2014

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1002/jbm.b.33162

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  • Construction of a bFGF-tethered multi-functional extracellular matrix protein through coiled-coil structures for neurite outgrowth induction

    Mie, Masayasu, Sasaki, Shoichi, Kobatake, Eiry

    Biomedical Materials   9 ( 1 )   2014

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1088/1748-6041/9/1/015004

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  • DNA-based immunoassays for sensitive detection of protein

    Akter, Farhima, Mie, Masayasu, Kobatake, Eiry

    Sensors and Actuators B-Chemical   202   2014

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1016/j.snb.2014.05.135

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  • Systematic screening of the cellular uptake of designed alpha-helix peptides Reviewed

    Usui, Kenji, Kikuchi, Takuya, Mie, Masayasu, Kobatake, Eiry, Mihara, Hisakazu

    Bioorganic & Medicinal Chemistry   21 ( 9 )   2560 - 2567   2013.5

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    DOI： 10.1016/j.bmc.2013.02.030

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  • A peptide release system using a photo-cleavable linker in a cell array format for cell-toxicity analysis Reviewed

    Kakiyama, Takashi, Usui, Kenji, Tomizaki, Kin-ya, Mie, Masayasu, Kobatake, Eiry, Mihara, Hisakazu

    Polymer Journal   45 ( 5 )   535 - 539   2013.5

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    DOI： 10.1038/pj.2013.20

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  • The promotion of angiogenesis by growth factors integrated with ECM proteins through coiled-coil structures

    Assal, Yasmine, Mie, Masayasu, Kobatake, Eiry

    Biomaterials   34 ( 13 )   2013

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1016/j.biomaterials.2013.01.067

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  • Development of a specific siRNA delivery system into HeLa cells using an IgG-binding fusion protein

    Bae, JooYoun, Mie, Masayasu, Kobatake, Eiry

    Biotechnology Letters   35 ( 12 )   2013

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    DOI： 10.1007/s10529-013-1299-y

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  • Hydrogel scaffolds composed of genetically synthesized self-assembling peptides for three-dimensional cell culture

    Mie, Masayasu, Oomuro, Mayu, Kobatake, Eiry

    Polymer Journal   45 ( 5 )   504 - 508   2013

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    DOI： 10.1038/pj.2012.216

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  • Selection of DNA Aptamers with Affinity for Pro-gastrin-Releasing Peptide (proGRP), a Tumor Marker for Small Cell Lung Cancer

    Mie, Masayasu, Kai, Tetsuro, Le, Thao, Cass, Anthony E. G., Kobatake, Eiry

    Applied Biochemistry and Biotechnology   169 ( 1 )   2013

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1007/s12010-012-9956-5

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  • Construction of multi-functional extracellular matrix proteins that inhibits migration and tube formation of endothelial cells Reviewed

    Nakamura, Makiko, Mie, Masayasu, Nakamura, Makoto, Kobatake, Eiry

    Biotechnology Letters   34 ( 8 )   1571 - 1577   2012.8

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    DOI： 10.1007/s10529-011-0788-0

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  • Targeted Gene Delivery via PEI Complexed with an Antibody

    Bae, JooYoun, Mie, Masayasu, Kobatake, Eiry

    Applied Biochemistry and Biotechnology   168 ( 8 )   2012

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1007/s12010-012-9928-9

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  • Construction of Semisynthetic DNA-Protein Conjugates with Phi X174 Gene-A* Protein

    Mashimo, Yasumasa, Maeda, Hitomi, Mie, Masayasu, Kobatake, Eiry

    Bioconjugate Chemistry   23 ( 6 )   2012

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    DOI： 10.1021/bc300118m

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  • Detection of Antigens Using a Protein-DNA Chimera Developed by Enzymatic Covalent Bonding with phiX Gene A

    Akter, Farhima, Mie, Masayasu, Grimm, Sebastian, Nygren, Per-Ake, Kobatake, Eiry

    Analytical Chemistry   84 ( 11 )   2012

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1021/ac300708r

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  • Development of a homogeneous immunoassay system using protein A fusion fragmented Renilla luciferase

    Mie, Masayasu, Ngo Phan Bich Thuy, Kobatake, Eiry

    Analyst   137 ( 5 )   2012

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1039/c2an15976g

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  • Development of a split SNAP-tag protein complementation assay for visualization of protein-protein interactions in living cells

    Mie, Masayasu, Naoki, Tatsuhiko, Uchida, Kentaro, Kobatake, Eiry

    Analyst   137 ( 20 )   2012

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1039/c2an35762c

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  • Aptamer-based protein detection using a bioluminescent fusion protein

    Akter, Farhima, Mie, Masayasu, Kobatake, Eiry

    Analyst   137 ( 22 )   2012

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1039/c2an35596e

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  • Construction of ECM-growth factor integrated protein through coiled-coil structure to promote angiogenesis

    Assal, Y., Mie, M., Kobatake, E.

    Journal of Tissue Engineering and Regenerative Medicine   6   299 - 299   2012

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  • Construction of affinity changeable antibody in response to Ca2+

    Kobatake, Eiry, Kosaku, Chihiro, Hanzawa, Satoshi, Mie, Masayasu

    Biotechnology Letters   34 ( 6 )   2012

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1007/s10529-012-0881-z

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  • Induction of motor neuron differentiation by transduction of Olig2 protein

    Mie, Masayasu, Kaneko, Mami, Henmi, Fumiaki, Kobatake, Eiry

    Biochemical and Biophysical Research Communications   427 ( 3 )   2012

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1016/j.bbrc.2012.09.090

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  • Cell fingerprint patterns using designed alpha-helical peptides to screen for cell-specific toxicity Reviewed

    Usui, Kenji, Kakiyama, Takashi, Tomizaki, Kin-ya, Mie, Masayasu, Kobatake, Eiry, Mihara, Hisakazu

    Bioorganic & Medicinal Chemistry Letters   21 ( 21 )   6281 - 6284   2011.11

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    DOI： 10.1016/j.bmcl.2011.09.002

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  • Targeted Delivery using Immunoliposomes with a Lipid-Modified Antibody-Binding Protein

    Kobatake, Eiry, Yamano, Ryo, Mie, Masayasu

    Applied Biochemistry and Biotechnology   163 ( 2 )   2011

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1007/s12010-010-9038-5

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  • Construction of a bFGF-Tethered Extracellular Matrix Using a Coiled-Coil Helical Interaction

    Kobatake, Eiry, Takahashi, Ryota, Mie, Masayasu

    Bioconjugate Chemistry   22 ( 10 )   2011

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1021/bc200249u

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  • Detection of small RNA molecules by a combination of branched rolling circle amplification and bioluminescent pyrophosphate assay

    Mashimo, Yasumasa, Mie, Masayasu, Suzuki, Shigeya, Kobatake, Eiry

    Analytical and Bioanalytical Chemistry   401 ( 1 )   2011

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1007/s00216-011-5083-3

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  • Development of an RNA detection system using bioluminescence resonance energy transfer

    Andou, Takashi, Endoh, Tamaki, Mie, Masayasu, Kobatake, Eiry

    Sensors and Actuators B-Chemical   152 ( 2 )   2011

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1016/j.snb.2010.12.020

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  • Direct detection of RNAs in living cells using peptide-inserted Renilla luciferase

    Andou, Takashi, Endoh, Tamaki, Mie, Masayasu, Kobatake, Eiry

    Analyst   136 ( 12 )   2011

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1039/c1an15130d

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  • Construction of a functional IgG-binding luciferase fusion protein for the rapid detection of specific bacterial strains Reviewed

    Nakamura, Makiko, Mie, Masayasu, Kobatake, Eiry

    Analyst   136 ( 1 )   71 - 72   2011

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    DOI： 10.1039/c0an00460j

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  • Immuno-rolling circle amplification using a multibinding fusion protein

    Akter, Farhima, Mie, Masayasu, Kobatake, Eiry

    Analytical Biochemistry   416 ( 2 )   2011

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1016/j.ab.2011.05.004

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  • Selection of DNA aptamers recognizing small cell lung cancer using living cell-SELEX

    Kunii, Takao, Ogura, Shun-ichiro, Mie, Masayasu, Kobatake, Eiry

    Analyst   136 ( 7 )   2011

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    DOI： 10.1039/c0an00962h

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  • Evaluation of small ligand-protein interaction by ligation reaction with DNA-modified ligand Reviewed

    Sugita, Rie, Mie, Masayasu, Funabashi, Hisakage, Kobatake, Eiry

    Biotechnology Letters   32 ( 1 )   97 - 102   2010.1

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    DOI： 10.1007/s10529-009-0109-z

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  • Construction of a thermostable cell adhesion protein for reverse transfection

    Bae, JooYoun, Goto, Sayaka, Mie, Masayasu, Kobatake, Eiry

    Journal of Biotechnology   150 ( 3 )   2010

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1016/j.jbiotec.2010.09.960

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  • Development of Renilla luciferase complementation system for homogeneous assay

    Mie, Masayasu, Andou, Takashi, Thuy, Ngo Phan Bich, Endoh, Tamaki, Kobatake, Eiry

    Luminescence   25 ( 2 )   2010

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  • Evaluation of small ligand-protein interactions by using T7 RNA polymerase with DNA-modified ligand

    Mie, Masayasu, Sugita, Rie, Endoh, Tamaki, Kobatake, Eiry

    Analytical Biochemistry   405 ( 1 )   2010

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1016/j.ab.2010.06.011

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  • Fluorescent and luminescent fusion proteins for detection of amyloid beta peptide localization and aggregation

    Usui, K., Mie, M., Andou, T., Sugimoto, N., Mihara, H., Kobatake, E.

    Journal of Peptide Science   16   175 - 175   2010

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  • Construction of a Multi-Functional Extracellular Matrix Protein That Increases Number of N1E-115 Neuroblast Cells Having Neurites

    Nakamura, Makiko, Mie, Masayasu, Mihara, Hisakazu, Nakamura, Makoto, Kobatake, Eiry

    Journal of Biomedical Materials Research Part B-Applied Biomaterials   91B ( 1 )   425 - 432   2009.10

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    DOI： 10.1002/jbm.b.31418

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  • Promotion of Angiogenesis by an Artificial Extracellular Matrix Protein Containing the Laminin-1-Derived IKVAV Sequence

    Nakamura, Makiko, Yamaguchi, Kumiko, Mie, Masayasu, Nakamura, Makoto, Akita, Keiichi, Kobatake, Eiry

    Bioconjugate Chemistry   20 ( 9 )   1759 - 1764   2009.9

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    DOI： 10.1021/bc900126b

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  • Construction of multifunctional proteins for tissue engineering: Epidermal growth factor with collagen binding and cell adhesive activities

    Imen, Elloumi Hannachi, Nakamura, Makiko, Mie, Masayasu, Kobatake, Eiry

    Journal of Biotechnology   139 ( 1 )   19 - 25   2009.1

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    DOI： 10.1016/j.jbiotec.2008.09.011

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  • Recombination System Based on Cre alpha Complementation and Leucine Zipper Fusions

    Seidi, Azadeh, Mie, Masayasu, Kobatake, Eiry

    Applied Biochemistry and Biotechnology   158 ( 2 )   2009

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1007/s12010-008-8409-7

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  • RNA detection using peptide-inserted Renilla luciferase

    Andou, Takashi, Endoh, Tamaki, Mie, Masayasu, Kobatake, Eiry

    Analytical and Bioanalytical Chemistry   393 ( 2 )   2009

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1007/s00216-008-2473-2

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  • Transduction of MyoD protein into myoblasts induces myogenic differentiation without addition of protein transduction domain

    Noda, Tomohide, Fujino, Takeshi, Mie, Masayasu, Kobatake, Eiry

    Biochemical and Biophysical Research Communications   382 ( 2 )   2009

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1016/j.bbrc.2009.03.060

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  • Construction of nanoscale protein particle using temperature-sensitive elastin-like peptide and polyaspartic acid chain

    Fujita, Yoshihiko, Mie, Masayasu, Kobatake, Eiry

    Biomaterials   30 ( 20 )   2009

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1016/j.biomaterials.2009.03.012

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  • Detection of Bioactive Small Molecules by Fluorescent Resonance Energy Transfer (FRET) in RNA-Protein Conjugates

    Endoh, Tamaki, Shintani, Ryo, Mie, Masayasu, Kobatake, Eiry, Ohtsuki, Takashi, Sisido, Masahiko

    Bioconjugate Chemistry   20 ( 12 )   2242 - 2246   2009

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    DOI： 10.1021/bc9002184

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  • Novel extracellular matrix for cell sheet recovery using genetically engineered elastin-like protein

    Mie, Masayasu, Mizushima, Yasunori, Kobatake, Eiry

    Journal of Biomedical Materials Research Part B-Applied Biomaterials   86B ( 1 )   2008

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1002/jbm.b.31019

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  • Selection of mRNA 5 '-untranslated region sequence with high translation efficiency through ribosome display

    Mie, Masayasu, Shimizu, Shun, Takahashi, Fumio, Kobatake, Eiry

    Biochemical and Biophysical Research Communications   373 ( 1 )   2008

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1016/j.bbrc.2008.05.173

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  • Combining electrical stimulation and cisplatin treatment increases caspase activity

    Mie, Masayasu, Manabe, Masafumi, Kobatake, Eiry

    Electrochemistry   76 ( 8 )   529 - 531   2008

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    DOI： 10.5796/electrochemistry.76.529

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  • Direct detection of RNA transcription by FRET imaging using fluorescent protein probe

    Endoh, Thmaki, Mie, Masayasu, Kobatake, Eiry

    Journal of Biotechnology   133 ( 4 )   2008

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1016/j.jbiotec.2007.11.005

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  • Assessment of small ligand-protein interactions by electrophoretic mobility shift assay using DNA-modified ligand as a sensing probe Reviewed

    Funabashi, Hisakage, Ubukata, Michito, Ebihara, Takashi, Aizawa, Masuo, Mie, Masayasu, Kobatake, Eiry

    Biotechnology Letters   29 ( 5 )   785 - 789   2007

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1007/s10529-006-9301-6

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  • Construction of intramolecular luciferase complementation probe for detecting specific RNA Reviewed

    Endoh, Tamaki, Mie, Masayasu, Funabashi, Hisakage, Sawasaki, Tatsuya, Endo, Yaeta, Kobatake, Eiry

    Bioconjugate Chemistry   18 ( 3 )   956 - 962   2007

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1021/bc060351o

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  • Design of a thermocontrollable protein complex

    Fujita, Yoshihiko, Funabashi, Hisakage, Mie, Masayasu, Kobatake, Eiry

    Bioconjugate Chemistry   18 ( 5 )   2007

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1021/bc070120x

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  • Novel recombination system using Cre recombinase alpha complementation

    Seidi, Azadeh, Mie, Masayasu, Kobatake, Eiry

    Biotechnology Letters   29 ( 9 )   2007

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1007/s10529-007-9406-6

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  • Detection of specific RNA by conformational change of recombinant protein

    Kobatake, Eiry, Endoh, Tamaki, Mie, Masayasu, Zhou, G, Lu, Z, Takeyama, H

    Progress on Post-Genome Technologies   2006

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  • Non-destructive monitoring of rpoS promoter activity as stress marker for evaluating cellular physiological status Reviewed

    Funabashi, H, Haruyama, T, Mie, M, Yanagida, Y, Kobatake, E, Aizawa, M

    Journal of Biotechnology   95 ( 1 )   85 - 93   2002

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    Publishing type：Research paper (scientific journal)  

    DOI： 10.1016/S0168-1656(01)00446-1

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  • Electrically enhanced differentiation of bone marrow stromal stem cells

    Mie, M, Ohgushi, H, Haruyama, T, Kobatake, E, Aizawa, M, Boom, H, Robinson, C, Rutten, W, Neuman, M, Wijkstra, H

    Proceedings of the 18th Annual International Conference of the Ieee Engineering in Medicine and Biology Society, Vol 18, Pts 1-5   18   1997

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Books

MISC

  • 抗血栓性人工血管の開発を目的とした細胞外マトリックスタンパク質の構築

    西岡宣之, 扇澤敏明, 三重正和, 小畠英理

    日本化学会講演予稿集   94th ( 3 )   2014

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  • Designed Short Peptides that Form Amyloid-Like Fibrils in Coassembly with Amyloid beta-Peptide (A beta) Decrease the Toxicity of A beta to Neuronal PC12 Cells

    Suzuki, Miho, Takahashi, Tsuyoshi, Sato, Junichi, Mie, Masayasu, Kobatake, Eiry, Mihara, Hisakazu

    Chembiochem   11 ( 11 )   1525 - 1530   2010

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  • Construction of multi-functional extracellular matrix proteins that promote tube formation of endothelial cells

    Makiko Nakamura, Masayasu Mie, Hisakazu Mihara, Makoto Nakamura, Eiry Kobatake

    BIOMATERIALS   29 ( 20 )   2977 - 2986   2008.7

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  • Construction of multi-functional extracellular matrix proteins that promote tube formation of endothelial cells

    Nakamura, Makiko, Mie, Masayasu, Mihara, Hisakazu, Nakamura, Makoto, Kobatake, Eiry

    Biomaterials   29 ( 20 )   2977 - 2986   2008.7

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  • Design of a Multifunctional Fusion Protein for Tissue Engineering

    ELLOUMI Imen, KOBAYASHI Rie, FUNABASHI Hisakage, MIE Masayasu, KOBATAKE Eiry

    2006   212 - 213   2007.3

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  • Bioluminescent enumeration of surface antigen-specific cells using the streptavidin-luciferase fusion protein

    Funabashi, Hisakage, Matsuzawa, Riko, Nakamura, Makiko, Mie, Masayasu, Kobatake, Eiry

    Sensors and Actuators B-Chemical   120 ( 1 )   51 - 56   2006.12

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  • Bioluminescent enumeration of surface antigen-specific cells using the streptavidin-luciferase fusion protein

    Hisakage Funabashi, Riko Matsuzawa, Makiko Nakamura, Masayasu Mie, Eiry Kobatake

    SENSORS AND ACTUATORS B-CHEMICAL   120 ( 1 )   51 - 56   2006.12

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  • Transduction of NeuroD2 protein induced neural cell differentiation

    Noda, Tomohide, Kawamura, Ryuzo, Funabashi, Hisakage, Mie, Masayasu, Kobatake, Eiry

    Journal of Biotechnology   126 ( 2 )   230 - 236   2006.11

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  • Transduction of NeuroD2 protein induced neural cell differentiation

    Tomohide Noda, Ryuzo Kawamura, Hisakage Funabashi, Masayasu Mie, Eiry Kobatake

    JOURNAL OF BIOTECHNOLOGY   126 ( 2 )   230 - 236   2006.11

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  • Delivery of antibody-captured proteins into living cells using PTD-fused protein A

    Masayasu Mie, Kazuto Mori, Hisakage Funabashi, Eiry Kobatake

    BIOTECHNOLOGY LETTERS   28 ( 15 )   1209 - 1214   2006.8

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  • Delivery of antibody-captured proteins into living cells using PTD-fused protein A

    Mie, Masayasu, Mori, Kazuto, Funabashi, Hisakage, Kobatake, Eiry

    Biotechnology Letters   28 ( 15 )   1209 - 1214   2006.8

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  • In vitro selection of zinc finger DNA-binding proteins through ribosome display

    Ihara, H, Mie, M, Funabashi, H, Takahashi, F, Sawasaki, T, Endo, Y, Kobatake, E

    Biochemical and Biophysical Research Communications   345 ( 3 )   1149 - 1154   2006.7

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  • In vitro selection of zinc finger DNA-binding proteins through ribosome display

    H Ihara, M Mie, H Funabashi, F Takahashi, T Sawasaki, Y Endo, E Kobatake

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   345 ( 3 )   1149 - 1154   2006.7

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  • Construction of epidermal growth factor fusion protein with cell adhesive activity

    Elloumi, I, R Kobayashi, H Funabashi, M Mie, E Kobatake

    BIOMATERIALS   27 ( 18 )   3451 - 3458   2006.6

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  • Construction of epidermal growth factor fusion protein with cell adhesive activity

    Elloumi, I, Kobayashi, R, Funabashi, H, Mie, M, Kobatake, E

    Biomaterials   27 ( 18 )   3451 - 3458   2006.6

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  • Cell-surface-localized ATP detection with immobilized firefly luciferase

    M Nakamura, M Mie, H Funabashi, K Yamamoto, J Ando, E Kobatake

    ANALYTICAL BIOCHEMISTRY   352 ( 1 )   61 - 67   2006.5

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  • Cell-surface-localized ATP detection with immobilized firefly luciferase

    M Nakamura, M Mie, H Funabashi, K Yamamoto, J Ando, E Kobatake

    ANALYTICAL BIOCHEMISTRY   352 ( 1 )   61 - 67   2006.5

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  • Fabrication of an antibody microwell array with self-adhering antibody binding protein

    Tanaka, G, Funabashi, H, Mie, M, Kobatake, E

    Analytical Biochemistry   350 ( 2 )   298 - 303   2006.3

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  • Fabrication of an antibody microwell array with self-adhering antibody binding protein

    G Tanaka, H Funabashi, M Mie, E Kobatake

    ANALYTICAL BIOCHEMISTRY   350 ( 2 )   298 - 303   2006.3

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  • Glucose oxidase assisted homogeneous electrochemical receptor binding assay for drug screening

    Funabashi, H, Tanaka, Y, Imamura, Y, Mie, M, Manabe, T, Tanaka, H, Takahashi, T, Handa, H, Aizawa, M, Kobatake, E

    Biosensors & Bioelectronics   21 ( 9 )   1675 - 1683   2006.3

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  • Glucose oxidase assisted homogeneous electrochemical receptor binding assay for drug screening

    H Funabashi, Y Tanaka, Y Imamura, M Mie, T Manabe, H Tanaka, T Takahashi, H Handa, M Aizawa, E Kobatake

    BIOSENSORS & BIOELECTRONICS   21 ( 9 )   1675 - 1683   2006.3

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  • Activity-based in vitro selection of T4 DNA ligase

    Takahashi, F, Funabashi, H, Mie, M, Endo, Y, Sawasaki, T, Aizawa, M, Kobatake, E

    Biochemical and Biophysical Research Communications   336 ( 3 )   987 - 993   2005.10

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  • Activity-based in vitro selection of T4 DNA ligase

    F Takahashi, H Funabashi, M Mie, Y Endo, T Sawasaki, M Aizawa, E Kobatake

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   336 ( 3 )   987 - 993   2005.10

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  • Method for detection of specific nucleic acids by recombinant protein with fluorescent resonance energy transfer

    Tamaki Endoh, Hisakage Funabashi, Masayasu Mie, Eiry Kobatake

    Analytical Chemistry   77 ( 14 )   4308 - 4314   2005.7

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  • Method for detection of specific nucleic acids by recombinant protein with fluorescent resonance energy transfer

    Endoh, T, Funabashi, H, Mie, M, Kobatake, E

    Analytical Chemistry   77 ( 14 )   4308 - 4314   2005.7

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  • Electrochemical evaluation of cellular physiological status under stress in Escherichia coli with the rpoS-lacZ reporter gene

    Hisakage Funabashi, Miyuki Ishikawa, Masayasu Mie, Fumio Takahashi, Yasuko Yanagida, Masuo Aizawa, Eiry Kobatake

    Biotechnology and Bioengineering   90 ( 4 )   509 - 515   2005.5

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  • Electrochemical evaluation of cellular physiological status under stress in Escherichia coli with the rpoS-lacZ reporter gene

    H Funabashi, M Ishikawa, M Mie, F Takahashi, Y Yanagida, M Aizawa, E Kobatake

    BIOTECHNOLOGY AND BIOENGINEERING   90 ( 4 )   509 - 515   2005.5

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  • Electrochemical evaluation of cellular physiological status under stress in Escherichia coli with the rpoS-lacZ reporter gene

    Funabashi, H, Ishikawa, M, Mie, M, Takahashi, F, Yanagida, Y, Aizawa, M, Kobatake, E

    Biotechnology and Bioengineering   90 ( 4 )   509 - 515   2005.5

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  • Construction of streptavidin-luciferase fusion protein for ATP sensing with fixed form

    Nakamura, M, Mie, M, Funabashi, H, Kobatake, E

    Biotechnology Letters   26 ( 13 )   1061 - 1066   2004.7

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  • On-chip biosensing of estrogen receptor-alpha at single molecular level

    DHB Wicaksono, T Ebihara, H Funabashi, M Mie, Y Yanagida, M Aizawa, E Kobatake

    BIOSENSORS & BIOELECTRONICS   19 ( 12 )   1573 - 1579   2004.7

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  • Construction of streptavidin-luciferase fusion protein for ATP sensing with fixed form

    M Nakamura, M Mie, H Funabashi, E Kobatake

    BIOTECHNOLOGY LETTERS   26 ( 13 )   1061 - 1066   2004.7

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  • On-chip biosensing of estrogen receptor-alpha at single molecular level

    Wicaksono, DHB, Ebihara, T, Funabashi, H, Mie, M, Yanagida, Y, Aizawa, M, Kobatake, E

    Biosensors & Bioelectronics   19 ( 12 )   1573 - 1579   2004.7

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  • Combined effect of electrical stimulation and cisplatin in HeLa cell death

    M Manabe, M Mie, Y Yanagida, M Aizawa, E Kobatake

    BIOTECHNOLOGY AND BIOENGINEERING   86 ( 6 )   661 - 666   2004.6

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  • Combined effect of electrical stimulation and cisplatin in HeLa cell death

    Manabe, M, Mie, M, Yanagida, Y, Aizawa, M, Kobatake, E

    Biotechnology and Bioengineering   86 ( 6 )   661 - 666   2004.6

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  • The construction of endothelial cellular biosensing system for the control of blood pressure drugs

    K Kamei, T Haruyama, M Mie, Y Yanagida, M Aizawa, E Kobatake

    BIOSENSORS & BIOELECTRONICS   19 ( 9 )   1121 - 1124   2004.4

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  • Construction and use of an electrochemical NO sensor in a cell-based assessing system

    Kamei, K, Mie, M, Yanagida, Y, Aizawa, M, Kobatake, E

    Sensors and Actuators B-Chemical   99 ( 1 )   106 - 112   2004.4

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  • The construction of endothelial cellular biosensing system for the control of blood pressure drugs

    Kamei, K, Haruyama, T, Mie, M, Yanagida, Y, Aizawa, M, Kobatake, E

    Biosensors & Bioelectronics   19 ( 9 )   1121 - 1124   2004.4

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  • Construction and use of an electrochemical NO sensor in a cell-based assessing system

    K Kamei, M Mie, Y Yanagida, M Aizawa, E Kobatake

    SENSORS AND ACTUATORS B-CHEMICAL   99 ( 1 )   106 - 112   2004.4

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  • Design of functional protein materials self-assembled on solid-phase surface

    E Kobatake, M Mie, M Aizawa

    MATERIALS TECHNOLOGY   19 ( 1 )   27 - 32   2004.3

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    Web of Science

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  • Design of functional protein materials self-assembled on solid-phase surface

    Kobatake, E, Mie, M, Aizawa, M

    Materials Technology   19 ( 1 )   27 - 32   2004.3

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  • 2A09-2 Development of detection method for specific nucleic acids by novel fusion protein

    ENDOH Tamaki, FUNABASHI Hisakage, MIE Masayasu, KOBATAKE Eiry

    16   103 - 103   2004

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    CiNii Books

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  • Electrically Modulated Cellular Biodevices

    Yasuko Yanagida, Masayasu Mie Eiry Kobatake, Masuo Aizawa

    Int.J.Soc.Mater.Eng.Resour.   12 ( 1 )   12 - 15   2004

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  • Electrically Modulated Cellular Biodevices

    Yasuko Yanagida, Masayasu Mie Eiry Kobatake, Masuo Aizawa

    Int.J.Soc.Mater.Eng.Resour.   12 ( 1 )   12 - 15   2004

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  • Design and construction of glutamine binding proteins with a self-adhering capability to unmodified hydrophobic surfaces as reagentless fluorescence sensing devices

    Wada, A, Mie, M, Aizawa, M, Lahoud, P, Cass, AEG, Kobatake, E

    Journal of the American Chemical Society   125 ( 52 )   16228 - 16234   2003.12

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  • Design and construction of glutamine binding proteins with a self-adhering capability to unmodified hydrophobic surfaces as reagentless fluorescence sensing devices

    A Wada, M Mie, M Aizawa, P Lahoud, AEG Cass, E Kobatake

    JOURNAL OF THE AMERICAN CHEMICAL SOCIETY   125 ( 52 )   16228 - 16234   2003.12

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  • Intracellular delivery of antibodies using TAT fusion protein A

    Mie, M, Takahashi, F, Funabashi, H, Yanagida, Y, Aizawa, M, Kobatake, E

    Biochemical and Biophysical Research Communications   310 ( 3 )   730 - 734   2003.10

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  • Intracellular delivery of antibodies using TAT fusion protein A

    M Mie, F Takahashi, H Funabashi, Y Yanagida, M Aizawa, E Kobatake

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   310 ( 3 )   730 - 734   2003.10

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  • Development of immune cellular biosensing system for assessing chemicals on inducible nitric oxide synthase signaling activator

    Kamei, K, Haruyama, T, Mie, M, Yanagida, Y, Aizawa, M, Kobatake, E

    Analytical Biochemistry   320 ( 1 )   75 - 81   2003.9

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  • Development of immune cellular biosensing system for assessing chemicals on inducible nitric oxide synthase signaling activator

    K Kamei, T Haruyama, M Mie, Y Yanagida, M Aizawa, E Kobatake

    ANALYTICAL BIOCHEMISTRY   320 ( 1 )   75 - 81   2003.9

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  • The secretory response through electric stimulation of differentiated PC12 rat pheochromocytoma cells transfected with neuropeptide Y fused with enhanced green fluorescent protein

    Mizuno, A, Mie, M, Yanagida, Y, Aizawa, M, Kobatake, E

    Biotechnology Letters   25 ( 7 )   547 - 552   2003.4

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  • The secretory response through electric stimulation of differentiated PC12 rat pheochromocytoma cells transfected with neuropeptide Y fused with enhanced green fluorescent protein

    A Mizuno, M Mie, Y Yanagida, M Aizawa, E Kobatake

    BIOTECHNOLOGY LETTERS   25 ( 7 )   547 - 552   2003.4

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  • Induction of neural differentiation by electrically stimulated gene expression of NeuroD2

    Mie, M, Endoh, T, Yanagida, Y, Kobatake, E, Aizawa, M

    Journal of Biotechnology   100 ( 3 )   231 - 238   2003.2

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  • Cellular biosensing system for assessing immunomodulating effects on the inducible nitric oxide synthase (iNOS) cascade

    Kamei, K, Haruyama, T, Mie, M, Yanagida, Y, Kobatake, E, Aizawa, M

    Biotechnology Letters   25 ( 4 )   321 - 325   2003.2

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  • Induction of neural differentiation by electrically stimulated gene expression of NeuroD2

    M Mie, T Endoh, Y Yanagida, E Kobatake, M Aizawa

    JOURNAL OF BIOTECHNOLOGY   100 ( 3 )   231 - 238   2003.2

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  • Cellular biosensing system for assessing immunomodulating effects on the inducible nitric oxide synthase (iNOS) cascade

    Ken-Ichiro Kamei, Tetsuya Haruyama, Masayasu Mie, Yasuko Yanagida, Eiry Kobatake, Masuo Aizawa

    Biotechnology Letters   25 ( 4 )   321 - 325   2003.2

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  • Ribosome display for selection of active dihydrofolate reductase mutants using immobilized methotrexate on agarose beads

    F Takahashi, T Ebihara, M Mie, Y Yanagida, Y Endo, E Kobatake, M Aizawa

    FEBS LETTERS   514 ( 1 )   106 - 110   2002.3

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  • Ribosome display for selection of active dihydrofolate reductase mutants using immobilized methotrexate on agarose beads

    Takahashi, F, Ebihara, T, Mie, M, Yanagida, Y, Endo, Y, Kobatake, E, Aizawa, M

    Febs Letters   514 ( 1 )   106 - 110   2002.3

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  • Fluorescent monitoring of cellular physiological status depending on the accumulation of ppGpp

    Funabashi, H, Mie, M, Yanagida, Y, Kobatake, E, Aizawa, M

    Biotechnology Letters   24 ( 4 )   269 - 273   2002.2

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  • Assessment of blood pressure control drugs with cellular biosensing system.

    亀井謙一郎, 三重正和, 柳田保子, 小畠英理, 相沢益男

    日本化学会講演予稿集   81st ( 2 )   2002

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  • Fluorescent monitoring of cellular physiological status depending on the accumulation of ppGpp

    Hisakage Funabashi, Masayasu Mie, Yasuko Yanagida, Eiry Kobatake, Masuo Aizawa

    Biotechnology Letters   24 ( 4 )   269 - 273   2002

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  • Cellular Biosensing System for Assessing Chemicals with the Inducible Nitric Oxide Synthase Cascade (Proceedings of The 5Th East Asian Conference on Chemical Sensors: The 33RD Chemical Sensor Symposium)

    Proceedings of the Chemical Sensor Symposium   33   95 - 97   2001.12

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  • Osteogenesis coordinated in C3H10T1/2 cells by adipogenesis-dependent BMP-2 expression system

    M Mie, H Ohgushi, Y Yanagida, T Haruyama, E Kobatake, M Aizawa

    TISSUE ENGINEERING   6 ( 1 )   9 - 18   2000.2

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  • Osteogenesis coordinated in C3H10T1/2 cells by adipogenesis-dependent BMP-2 expression system

    Mie, M, Ohgushi, H, Yanagida, Y, Haruyama, T, Kobatake, E, Aizawa, M

    Tissue Engineering   6 ( 1 )   9 - 18   2000.2

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  • Electrically enhanced osteogenic differentiation of rat bone marrow stromal stem cells

    Masayasu Mie, Hajime Ohgushi Tetsuya, Haruyama Eiry Kobatake, Masuo Aizawa

    J. Cellular Engineering   1 ( 4 )   153 - 158   1996

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  • Electrically enhanced osteogenic differentiation of rat bone marrow stromal stem cells

    Masayasu Mie, Hajime Ohgushi Tetsuya, Haruyama Eiry Kobatake, Masuo Aizawa

    J. Cellular Engineering   1 ( 4 )   153 - 158   1996

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Presentations

  • 不凍タンパク質による非特異的なタンパク質吸着の抑制効果

    堀之内 悠真, 三重 正和, 小畠 英理, 西田 慶

    化学工学会 第90年会  2025.3 

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  • Protein Ligation を用いた高機能化タンパク質ナノ粒子の構築

    小椋 優菜, 山口 醇, 西田 慶, 小畠 英理, 三重 正和

    化学工学会 第90年会  2025.3 

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  • DNA の二重鎖形成に依存したSplit intein 活性回復を利用したタンパク質ライゲーション法の開発

    森下 祐衣, 西田 慶, 小畠 英理, 三重 正和

    化学工学会 第90年会  2025.3 

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  • 細胞膜コレステロール結合性リコンビナントタンパク質による細胞操作

    西田 慶, 三重 正和, 小畠 英理

    化学工学会 第90年会  2025.3 

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  • 膜界面間の相互作用を制御するssDNA・コレステロール結合性融合タンパク質の開発

    大場 奈海, 石塚 美音, 西田 慶, 小畠 英理, 三重 正和

    日本バイオマテリアル学会シンポジウム2024  2024.10 

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  • 不凍タンパク質の凍結保護性に対するELPの融合化が与える影響

    竹山 颯一, 西田 慶, 三重 正和, 小畠 英理

    第14回 CSJ化学フェスタ2024  2024.10 

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  • Modulation of cellular metabolism evoked by recombinant proteins binding to plasma membrane cholesterol

    Kei Nishida, masayasu mie, EIRY KOBATAKE

    2024.12 

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  • Evaluation of intracellular-delivered IgG by binding to cell-penetrating peptide via IgG-binding domain

    2024.12 

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  • ヒアルロン酸の検出と細胞機能の制御を目指したCD44融合分子の構築

    屋祢下創太, 西田 慶, 三重正和, 小畠英理

    第53回医用高分子シンポジウム  2024.7 

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  • タンパク質ナノ粒子の構築を指向したCatcher/Tagシステムによる機能性タンパク質環状化

    藤原 快, 西田 慶, 市川 真, 山口 醇, 三重 正和, 小畠 英理

    第76回 日本生物工学会大会  2024.9 

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  • 細胞遊走性を抑制する膜結合性タンパク質材料の構築

    西田 慶, 三重 正和, 小畠 英理

    2024.9 

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  • 翻訳後切断を指向したDNA結合タンパク質Repの改変

    村山 瑞季, 西田 慶, 小畠 英理, 三重 正和

    第18回バイオ関連化学シンポジウム  2024.9 

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  • 癌細胞の機能制御を目的とした膜コレステロール結合性タンパク質の合成と機能評価

    佐藤郁美, 西田 慶, 小畠英理, 三重正和

    第53回医用高分子シンポジウム  2024.7 

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  • 直鎖状融合タンパク質によるバイオ界面の設計とアンチファウリング特性の評価

    西田 慶, 三重正和, 小畠英理

    第53回医用高分子シンポジウム  2024.7 

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  • ssDNAおよびコレステロール結合性融合タンパク質による脂質膜表面の改質と機能評価

    大場 奈海, 石塚 美音, 西田 慶, 小畠 英理, 三重 正和

    第40回日本DDS学会学術集会  2024.7 

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  • 膜コレステロール結合性タンパク質の高分子修飾と機能評価

    馬天健, 西田慶, 佐藤郁美, 三重正和, 小畠英理

    日本化学会 第104春季年会  2024.3 

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  • Catcher/Tagシステムを利用した環状化Renilla Luciferaseの構築

    市川真, 西田慶, 山口醇, 小畠英理, 三重正和

    日本化学会 第104春季年会  2024.3 

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  • Sensitive detection and capture of tumor cells using protein nanoparticles with multiple display of DNA aptamers and bioluminescent reporters International conference

    Kei Nishida, Gaoyang Wang, Masayasu Mie, Eiry Kobatake

    The 12th World Biomaterials Congress（WBC2024）  2024.5 

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  • Construction of multifunctional protein surface containing Elastin-like polypeptide for tissue engineering International conference

    Mutawakil Al Muqadasi, Kei Nishida, Masayasu Mie, Eiry Kobatake

    The 12th World Biomaterials Congress（WBC2024）  2024.5 

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  • Repを利用したタンパク質の直交型DNA修飾

    小松﨑千加, 西田慶, 小畠英理, 三重正和

    日本化学会 第104春季年会  2024.3 

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  • 電気化学ホモジニアスアッセイによるドラッグスクリーニング法の開発

    電気化学会第71回大会  2004 

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  • 電気刺激による神経突起伸長方向制御法の開発

    電気化学会第72回大会  2005 

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Awards

  • International Academy for Medical and Biological Engineering Prizes for young scientists, highly commended-best poster/paper

    2003  

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  • 鎌田泉博士論文賞

    2000  

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Research Projects

  • 融合タンパク質の網羅的構築を可能にするProtein Ligation法の開発

    Grant number：24K21695  2024.6 - 2026.3

    日本学術振興会  科学研究費助成事業  挑戦的研究(萌芽)

    三重 正和

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    Grant amount：\6500000 （ Direct Cost: \5000000 、 Indirect Cost：\1500000 ）

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  • シグナル因子提示細胞を用いた細胞機能プログラム可能なボトムアップ型組織構築法開発

    Grant number：23K28427  2023.4 - 2026.3

    日本学術振興会  科学研究費助成事業  基盤研究(B)

    三重 正和

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    Grant amount：\12220000 （ Direct Cost: \9400000 、 Indirect Cost：\2820000 ）

    本研究は、細胞間接着を基盤とした組織構築法に細胞の分化・増殖を制御するシグナル因子を導入した新規ボトムアップ型組織構築システムの開発を目的とする。このシステムでは、DNAとタンパク質を結合したハイブリッド分子を構築し、細胞表面に提示したDNAを介して細胞間接着、機能性タンパク質の提示を行う。ここではコレステロール結合能を有するタンパク質のドメイン4(ALOD4)と任意の一本鎖DNA(ssDNA)配列との結合能を有するブタサーコウイルス由来複製開始タンパク質(Rep)の融合タンパク質Rep-ALOD4を構築し、Repを介してDNA-ALOD4ハイブリッド分子を構築した。蛍光標識したDNAとALOD4のハイブリッド分子を細胞に添加したところ、細胞表面に沿って蛍光が観察された。このことから、細胞表面へのDNA提示が示された。このALOD4は、細胞膜中のコレステロールに結合した後は、細胞内に取り込まれることなく細胞膜から離脱する。そこで、細胞表面に提示されたDNAの細胞膜表面滞留性を向上させるためにALOD4をタンデムに融合したRep-(ALOD4)2を構築した。Rep-(ALOD4)2を用いても、Rep-ALOD4同様に細胞表面にDNAの提示は可能であった。また、その細胞膜表面滞留性が向上していることが示唆された。次に機能性タンパク質の細胞表面提示を目指し、モデルタンパク質としてGFPとRepの融合タンパク質(GFP-Rep)を構築した。Rep-ALOD4を介して細胞表面に提示したssDNAと相補的な配列を有するssDNAとGFP-Repのハイブリッドを、DNAを提示した細胞に添加したところ、DNAの二本鎖形成を介した細胞表面への機能性タンパク質の提示に成功した。

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  • Construction of Nanostructure with DNA-protein hybrid molecules for cell reprogramming

    Grant number：16K01388  2016.4 - 2020.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

    Mie Masayasu

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    Grant amount：\4680000 （ Direct Cost: \3600000 、 Indirect Cost：\1080000 ）

    In this research, a novel method for cell reprogramming by using DNA-protein hybrid molecules is developed. To induce cellular reprogramming, combinations of transcription factors should be introduced into cells. To form complex of transcription factors, DNA-protein hybrid molecules are focused. Firstly, to clarify whether combinations of protein can be introduced into a cell, transcription factor combined with Green fluorescence protein was introduced into cells. From the results, it was clarified that combination of protein which formed complex can be introduced into cells. Secondly, a new method for construction of DNA-protein hybrid molecule is developed. By fusing a protein of interest to replication initiator protein, DNA-protein hybrid molecules can be constructed.

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  • Construction of a tissue-specific transcription factor-tethered extracellular matrix protein

    Grant number：24650276  2012.4 - 2015.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Challenging Exploratory Research

    MIE Masayasu

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    Grant amount：\3900000 （ Direct Cost: \3000000 、 Indirect Cost：\900000 ）

    In this study, a novel strategy for the construction of biomaterials is introduced by tethering a tissue-specific transcription factor protein to an artificial extracellular matrix (ECM) protein. Here, oligodendrocyte and motor neuron specific transcription factor Olig2 was tethered to an artificial ECM protein via coiled-coil helix formation. The artificial ECM, is comprised of an elastin-like peptide as a structural unit, as well as the AG73 peptide sequence derived from laminin and the C3 peptide sequence which binds to neural cell adhesion molecules (NCAMs) derived from the synthetic peptide library, for cell adhesion and enhance neurite outgrowth. As a proof of concept, tethered Olig2 was internalized into mouse embryonic carcinoma P19 cells and the ability to induce neural differentiation was investigated.

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  • Development of a novel active targeting system based on split SNAP-tag

    Grant number：22680041  2010.4 - 2013.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Young Scientists (A)

    MIE Masayasu

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    Grant amount：\21320000 （ Direct Cost: \16400000 、 Indirect Cost：\4920000 ）

    In this experiment, a novel active targeting system based on split SNAP-tag was developed. Firstly, split SNAP-tag complementation was studied. Split SNAP-tags, fragments of divided SNAP-tag were fused to proteins that can interact with each other. After incubation with a fluorescent SNAP-tag substrate, cells that expressed split SNAP-tag fusion proteins generated fluorescent signals when these proteins interacted. It was shown that split SNAP-tag labeling method should be a useful tool for visualization of protein-protein interaction processes.
    Next, for using DNA aptamer in our system, we developed a method for site-specific labeling of single-stranded DNA (ssDNA) to a recombinant protein of interest (POI) through the Gene-A* protein (Gene-A*) from bacteriophage phi X174, without any chemical modifications of ssDNA.

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  • Construction of cellular imaging system by delivery of antibody into living cells

    Grant number：20700376  2008 - 2009

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Young Scientists (B)

    MIE Masayasu

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    Grant amount：\4160000 （ Direct Cost: \3200000 、 Indirect Cost：\960000 ）

    The purpose of this experiment was construction of imaging system in living cell by delivery of antibody. We have constructed imaging system which shows the signal when antibody bind to a target molecule. It was showed that our imaging system would be applied for living cell imaging.

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  • Construction of Intelligent Cellular Biosensing System

    Grant number：16360407  2004 - 2006

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)

    KOBATAKE Eiry, MIE Masayasu

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    Grant amount：\14000000 （ Direct Cost: \14000000 ）

    Cells can growth and differentiate by the effect of some growth factors. Cells also have various intelligent systems such as self-diagnosis and self-repair. Therefore, we have a possibility to realize advanced intelligent biosensing systems through the use living cells as materials. In this study, we purposed to construct an intelligent cellular biosensing system which can evaluate the effects of chemicals to a living body.
    We had already established a cellular biosensing system for assessing chemicals which have the effect to the endothelial cells by using NO released from cells. In this year, we tried to construct a novel cellular biosensing system for the detection of allergens. For this purpose, we utilized a histamine oxidase. At first, histamine oxidase was expressed in E.coli and purified by affinity chromatography. Some electrochemical characterizations were then performed. Finally, we constructed a biosensing system by the combination of cells and histamine oxidase-immobilized electrode. We could detect histamine which was released from cells by the stimulation of chemicals.
    From these results, it is clarified that constructing intelligent cellular biosensing system is useful to evaluate the effects of chemicals to a living body.

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  • "Deliver-body"システムによるタンパク質の細胞内導入

    Grant number：16700353  2004 - 2006

    日本学術振興会  科学研究費助成事業  若手研究(B)

    三重 正和

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    Grant amount：\3000000 （ Direct Cost: \3000000 ）

    本研究では、細胞膜透過能を有するTAT-PTDと抗体結合能を有するプロテインAのBドメインを融合したタンパク質(TAT-B2C)による抗体の細胞内への導入技術を基に、新たなタンパク質の細胞内導入法の開発を目的とする。
    前年度、神経細胞分化誘導を目的としてdeliver-bodyを用いて細胞内に導入するために精製を行った転写因子NeuroD2タンパク質が膜透過能を有する事を見出した。本年度は、この転写因子NeuroD2の細胞膜透過能、並びにその分化誘導能について検討を行った。
    始めにNeuroD2内に存在する膜透過配列を明らかにするためNeuroD2内の塩基性に富んだ配列を検索し、その配列を欠損したNeuroD2タンパク質を作製した。作製したタンパク質を蛍光標識し細胞に添加したところ、100からIl5番目に存在する核移行シグナルが膜透過能に関与している事が示唆された。更にTAT-PTDの代わりにプロテインAのBドメインにこの配列を融合し、その膜透過能を検討したところTAT-PTD同様に膜透過能を有することが明らかになった。次に導入されたNeuroD2タンパク質が神経細胞分化を誘導可能であるかを細胞の形態変化とRT-PCRによる遺伝子発現解析により検討した。その結果、細胞内に導入されたNeuroD2タンパク質が神経細胞分化を誘導することを明らかにした。
    これらの結果から、転写因子NeuroD2はdeliver bodyシステムを用いなくても細胞内に導入することが可能であり、更には細胞分化をも誘導可能であることが明らかになった。

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  • 骨組織再生を目的とした細胞間情報ネットワークの創製

    Grant number：99J06492  1999 - 2000

    日本学術振興会  科学研究費助成事業  特別研究員奨励費

    三重 正和

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    Authorship：Principal investigator 

    Grant amount：\1800000 （ Direct Cost: \1800000 ）

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