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Rapid and low cost synthesis of monoclonal antibody from single B cells of blood.

  • School of Agricultural Science/Graduate School of Bioagricultural Sciences
  • Laboratory of Molecular Biotechnology

Hideo Nakano [Professor]

http://www.agr.nagoya-u.ac.jp/~molbiote/index-e.html

Outline of Seeds

Monoclonal antibody (Mab) has been used in numerous fields including clinical examinations, medicines, biosensors, food safety control and so on. But conventional methods to produce Mab such as hybridoma method and phage display have several drawbacks in terms of cost and time involved in obtaining and producing Mab. In addition, animal species suitable for Mab production are limited. Our research group, however, has come up with a technology that allows us to amplify the cDNA of antibody in just two days from single B cells displaying antibodies binding to a target molecule and rapidly synthesize Fab antibodies using cell-free protein synthesis. We have created a start-to-finish production system called the Ecobody Method, which makes it possible to screen Mab from single B cells and to produce enough amount of Fab antibodies by large-scale synthesis using E. coli (see figure).

Novelty and originality of this research

Compared to the hybridoma method, our method allows us to obtain monoclonal antibodies at extremely low cost and in a short period of time simply by making extensive use of molecular biological technologies such as PCR and cell-free protein synthesis. By selecting B cells that display antibodies in advance, it is easy to synthesize Fab that has the target selectivity and compatibility. Also, in addition to being able to obtain monoclonal antibodies independent of the target animal species, we can also rapidly synthesize human antibodies from B cells derived from trace amounts of human peripheral blood.

Application and research area for Industry collaboration

Support for drug development, analysis of human antibodies

Monographs, Papers and Articles

  • Ojima-Kato, T et al. (2016) "Zipbody" leucine zipper-fused Fab in E. coli in vitro and in vivo expression systems. Protein Eng. Des. Sel. 29, 149-157
  • Ojima-Kato, T.et al. (2015) In vitro generation of rabbit anti- Listeria monocytogenes monoclonal antibody using single cell based RT-PCR linked cell-free expression systems. J. Immunol. Methods 427, 58-65.
  • Jiang, X.P. et al. (2006) A Novel Strategy for Generation of Monoclonal Antibodies from Single B Cells Using RT-PCR Technique and in Vitro Expression. Biotechnol. Prog., 22, 979-988.