Construction and Expression of Human Anti-Keratin ScFv
- VernacularTitle:人源性抗角蛋白单链抗体的构建及表达
- Author:
Chengxin LI
;
Yufeng LIU
;
Gang WANG
;
Chunying LI
;
Yehong WAN
;
Wei LI
- Publication Type:Journal Article
- Keywords:
Keratin;
Antibodies;
DNA,recombinant;
Genes
- From:
Chinese Journal of Dermatology
1995;0(04):-
- CountryChina
- Language:Chinese
-
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 i-dentified by ELISA.Results Results of DNA sequence analysis showed that nucleotide sequence of V?and VH of ScFv gene was similar to that of V?and 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 an-ti-keratin ScFv might lay a solid foundation for further observation of biological activities of anti-keratin au-toantibody,manufacture of genetic-engineering anti-keratin products,and promotion of clinical application of genetic-engineering anti-keratin antibodies.
