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Generation of anti-c-met single domain antibody fragment based on human stable frameworks

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Abstract

The size reduction is an important issue in the biomedical application of antibody and single domain antibody fragment is recognized as very attractive tool. However, it is very time-consuming and laborious to generate the fragment antibody with targeted binding function. Here, we investigated the possibility to prepare single domain antibody (sdAb) by a simple grafting method based on stable human consensus framework sequences. The complementarity determining region sequences in VH domain of anti-c-Met scFv from rabbit were grafted with the human VH3 consensus framework sequences, which generated the anti-c-Met single domain antibody showing almost same binding activity to its scFv form. The generated single domain antibody could be produced as functional form in oxidizing cytoplasm of E. coli, but produced as inactive form in reducing cytoplasm. The structural analysis of the homology models gave us the insight on the stability of the single domain antibody. In this report, we have demonstrated that the very stable human consensus framework sequence can be used for the generation of active anti-c-Met sdAb via complementarity determining regions grafting. We expect that this kind of grafting method for the generation of sdAb may provide us with the opportunities to prepare sdAbs based on the known antibody sequences.

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Authors and Affiliations

  1. Department of Chemical Engineering, Pusan National University, Busan, 609-735, Korea

    Selvakumar Edwardraja, Sriram Sokalingam, Govindan Raghunathan & Sun-Gu Lee

  2. Department of Chemistry and Biochemistry, Arizona State University, Arizona, 85287, USA

    Selvakumar Edwardraja

  3. Department of Chemical and Biomolecular Engineering, QB3 Institute, University of California-Berkeley, Berkeley, CA, 94720, USA

    Bum-Yeol Hwang

Authors
  1. Selvakumar Edwardraja
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  2. Sriram Sokalingam
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  3. Govindan Raghunathan
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  4. Bum-Yeol Hwang
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  5. Sun-Gu Lee
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Correspondence to Bum-Yeol Hwang or Sun-Gu Lee.

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Edwardraja, S., Sokalingam, S., Raghunathan, G. et al. Generation of anti-c-met single domain antibody fragment based on human stable frameworks. Biotechnol Bioproc E 17, 1120–1127 (2012). https://doi.org/10.1007/s12257-012-0378-6

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  • DOI: https://doi.org/10.1007/s12257-012-0378-6

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