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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Expression Vectors for Human-mouse Chimeric Antibodies

  • Xiong, Hua (Cell Engineering Research Center, State Key Laboratory of Cancer Biology, Fourth Military Medical University) ;
  • Ran, Yuliang (Department of Cell and Molecular Laboratory, Cancer Institute (Hospital), Chinese Academy of Medical Sciences and Peking Union Medical College) ;
  • Xing, Jinliang (Cell Engineering Research Center, State Key Laboratory of Cancer Biology, Fourth Military Medical University) ;
  • Yang, Xiangmin (Cell Engineering Research Center, State Key Laboratory of Cancer Biology, Fourth Military Medical University) ;
  • Li, Yu (Cell Engineering Research Center, State Key Laboratory of Cancer Biology, Fourth Military Medical University) ;
  • Chen, Zhinan (Cell Engineering Research Center, State Key Laboratory of Cancer Biology, Fourth Military Medical University)
  • Published : 2005.07.31
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Abstract

The production of recombinant antibodies has been generally recognized as time-consuming and labor-intensive. The aim of our study is to construct mammalian expression vectors containing the cDNA encoding the human constant regions and murine variable regions to massively and cost-effectively produce full-length chimeric antibodies. Unique restriction sites flanking the Ig variable region were designed to allow for the replacement of variable regions generated by PCR. Western blot analysis of the chimeric antibodies revealed that the expressed products were of the predicted size, structure and specificity. The usefulness of the vectors was confirmed by construction of human-mouse chimeric antibody-HCAb which secretes murine antibody against the human colorectal cancer. Selected in medium containing gradually increasing methotrexate (MTX), clones with increased expression of the product gene can be efficiently generated. The secretion of recombinant chimeric antibody-HCAb yielded $30\;pg\;cell^{-1}\;day^{-1}$ at $10^{-6}\;M$ MTX. With this high-level expression from pools, the convenient and rapid production of over 100 milligram amounts per liter of recombinant antibodies may be achieved, which indicates the significant roles of pYR-GCEVH and pYR-GCEVL in the production of chimeric antibodies.

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References

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