Construction of Human Antibody Library and Analysis of its Diversity

TIAN Xue jun,SHOU Cheng chao

Chinese Journal of Biochemistry and Molecular Biology ›› 2000, Vol. 16 ›› Issue (02) : 200-205.

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PDF(360 KB)
Chinese Journal of Biochemistry and Molecular Biology ›› 2000, Vol. 16 ›› Issue (02) : 200-205.
Article

Construction of Human Antibody Library and Analysis of its Diversity

  • TIAN Xue jun,SHOU Cheng chao
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Abstract

Human antibody library was constructed by using phage display technology and phage Fab antibodies against vascular endothelial growth factor(VEGF)were screened from this library.Enzyme analysis of these Fab antibodies was accomplished for diversity.Human immunoglobulin heavy chain and light chain genes were separately amplified by RT PCR from human peripheral lymphocytes using family specific primers and signal sequences of immunoglobulin as half nested PCR.The heavy and light chain PCR products were then cloned into phagemid vector pComb3H and a repertoire of 1.5×10 8 clones of Fab fragments displayed on the surface of phage was constructed.After 4 rounds panning with VEGF 121 ,280 clones were checked for their binding activities with ELISA and 12 clones could bind to VEGF 121 specifically.Restriction enzyme analysis showed that these clones contained vast diversity.These data provided a new way for preparation of high affinity human anti VEGF monoclonal antibodies through antibody engineering.

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TIAN Xue jun,SHOU Cheng chao. Construction of Human Antibody Library and Analysis of its Diversity[J]. Chinese Journal of Biochemistry and Molecular Biology, 2000, 16(02): 200-205

References

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