Screening of Human Anti-α-crystallin Protein of Mycobacterium Tuberculosis scFv from Large Phage Library

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China Biotechnology ›› 2012, Vol. 32 ›› Issue (09) : 22-27.

WANG Xiao-na¹, MI Zhi-qiang¹, AN Xiao-ping¹, LI Jian-bin¹, FAN Hua-hao¹, ZHANG Wen-hui¹, ZHANG Bo¹, HUANG Yong¹, ZHOU Li-jun², TONG Yi-Gang ¹

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Abstract

Objetive:To obtain the specific human scFv against Mycobacterium tuberculosis Acr protein using phage antibody library technology.Methods: A human large phage antibody library was panned four times with the purified Acr protein.Soluble scFvs were prepared through infection of E.coli HB2151 with the selected phage antibodies clones and induction with IPTG. The antigen binding activity were determined ELISA and Western blotting, and DNA sequences of these clones were analyzed by Sanger sequencing.Results:43 positive clones were obtained after 4 rounds of panning and 29 clones had binding ability to Mycobacterium tuberculosis Acr protein. DNA sequencing of the 29 clones showed 26 different positive clones. 14 specific soluble scFvs were obtained after infection in E.coli HB2151. DNA sequence analysis showed that the variable regions of these scFvs belonged to different subgroups.The Western blotting results suggested that the anti-Acr antibodies had good immuno-reactivity with natural Acr protein.Conclusion:Human anti-Acr scFvs are obtained from a large phage antibody library, which will benefit to the application of anti-Acr of Mycobacterium tuberculosis antibody to clinical research.

Key words

Mycobacterium tuberculosis / Acr protein / Phage antibody library / scFv

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WANG Xiao-na, MI Zhi-qiang, AN Xiao-ping, LI Jian-bin, FAN Hua-hao, ZHANG Wen-hui, ZHANG Bo, HUANG Yong, ZHOU Li-jun, TONG Yi-Gang. Screening of Human Anti-α-crystallin Protein of Mycobacterium Tuberculosis scFv from Large Phage Library[J]. China Biotechnology, 2012, 32(09): 22-27

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