Chinese Journal of Dermatology ›› 2003, Vol. 36 ›› Issue (4): 181-184.

    Next Articles

Construction and Expression of Human Anti-Keratin ScFv

LI Cheng-xin, LIU Yu-feng, WANG Gang, LI Chun-ying, WAN Ye-hong, LI Wei   

  1. Department of Dermatology, Xijing Hospital, Fourth Military Medical University, Xi'an 710033, China
  • Received:2002-04-04 Online:2003-04-15 Published:2003-04-15

Abstract: Objective To construct and express human ScFv against keratin in E.coli,and identify its binding activity with antigens and antigenic specificity.Methods By genetic engineering technology,human anti-keratin ScFv was constructed from Fab fragment selected from established semi-synthetic phage antibody library.The binding activity with antigens and antigenic specificity of expressed products were identified by ELISA.Results Results of DNA sequence analysis showed that nucleotide sequence of Vκand VH of ScFv gene was similar to that of Vκand VH of Fab gene,suggesting that there was no mutation in the construction of ScFv.The soluble anti-keratin ScFv was found to have a good antigenic specificity as well as excellent binding activity with target antigens by ELISA.However,the binding activity of ScFv with keratin was lower than that of Fab.Conclusions The successful expression and identification of human anti-keratin ScFv might lay a solid foundation for further observation of biological activities of anti-keratin autoantibody,manufacture of genetic-engineering anti-keratin products,and promotion of clinical application of genetic-engineering anti-keratin antibodies.

Key words: Keratin, Antibodies, DNA,recombinant, Genes