Construction of a human phage display ScFv library against mycobacterium tuberculosis
- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF TUBERCULOSIS AND RESPIRATORY DISEASES
- Issue:
- 9
- DOI:
- 10.3321/j.issn:1001-0939.2008.09.014
- Key Word:
- 分枝杆菌,结核;噬菌体展示;单链抗体;文库;Mycobacteridum tuberculosis;Phage display;Single-chain Fv antibody;Library
Abstract： Objective To construct a human phage display single-chain Fv(ScFv)antibody library against mycobacterium tuberculosis(MTB),for specific ScFv antibody cloning.Methods Total RNA was isolated from the lymphocytes of patients with positive serum antibody against MTB and reveree transcribed into cDNA.The heavy chain and light chain variable region gene of human immunoglobulin were amplified individually by PCR and then assembled into ScFv genes.The ScFv genes were Hgated into phagemid pCANTAB5S.The human phage display ScFv library agaiast MTB was constructed by transforming the recombinant phagemid into E.coli TG1 with the presence of helper phage M13K07.Results The human phage display ScFv library containing 107 different clones was constructed successfully.Conclusions A phage display ScFv library against MTB has been constructed based on the variable region gene of immunoglobulin of the lymphocytes of TB patients and phagemid pCANTABSS.The specific ScFv antibodies can be screened from this library.
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