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| Construct and Characterization of Human Naive ScFv Phage Display Library |
| MAO Xiao-yan,QIAO Yu-ling,LU Wei-jia,MA Rui,ZHAO Hong |
| Lanzhou Institute of Biological Products, Lanzhou 730046, China
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Abstract Phage display antibody library is an important technology for generating therapeutic antibody.It was reported that the construction and characterization of a large human antibody phage display library. Total RNA was extracted from peripheral blood lymphocytes of 20 healthy donors. VH and VL gene were amplified by RT-PCR. The ScFv (single chain variable fragment) gene were assembled by overlap PCR, and cloned into a phagemid vector by electroporation of E.coli TG1. A large naive human ScFv phage antibody library containing 1.3×109 antibody members was constructed by repetitive electroporation about 300 times. The quality of the library was examined by sequence analysis and selection against 5 different protein antigens. Sequence analysis of the randomly picked 40 clones showed this library had good diversity. Panning of the 5 different antigens all resulted in successful isolation of antigen specific ScFv antibodies. The results indicate that a large nave human ScFv phage antibody library with good diversity was constructed successfully.
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Received: 08 December 2009
Published: 25 May 2010
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