[Construction and expression of anti-keratin single-chain Fv antibody].
OBJECTIVE
To construct an expression vector of human anti-keratin scFv and express functional scFv in E. coli.


METHODS
The V(H) and V(L) genes were amplified by PCR and cloned into a scFv expression vector, which was expressed in E. coli. The expression of scFv was tested by SDS-PAGE; The antigen-binding characteristics of scFv were tested by ELISA.


RESULTS
The V(H) and V(L) genes were cloned and expressed in E. coli successfully. The expressed scFv retained the antigen-binding characteristics of the parental antibody.


CONCLUSIONS
A functional anti-keratin scFv was obtained.
Zhang
Guo-min
GM
Department of Microbiology and Immunology, Capital University of Medical Science, Beijing 100054, China. erminzhang@sohu.com
Chen
Yu-ping
YP
Wang
Gang
G
Liu
Yu-feng
YF
D000906:Antibodies / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification / Q000494:pharmacology; D000918:Antibody Specificity / Q000235:genetics / Q000276:immunology*; D015153:Blotting, Western; D002460:Cell Line; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification / Q000494:pharmacology; D007633:Keratins / Q000276:immunology*; D019151:Peptide Library; D011485:Protein Binding
D000906:Antibodies; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D007633:Keratins