[Affinity maturation of a single-chain antibody against hepatocellular carcinoma].
OBJECTIVE
To obtain a single-chain antibody with high affinity against hepatocellular carcinoma (HCC).


METHODS
A second single-chain antibody mutant library was established using an error-prone PCR and a phage display. Single-chain antibodies with high affinity for hepatocellular carcinoma were selected using ELISA.


RESULTS
The content of the second single-chain antibody mutant library was about 4.5 x 10(7). Two selected mutants, M90 and M116, were obtained after 3 rounds of panning and ELISA. Immunoassay showed that both M90 and M116 could bind to human HCC cells. The relative affinity of M90 was 1.7-fold higher than that of the original antibody, and M116 was 2-fold higher than that of the original antibody.


CONCLUSIONS
Error-prone PCR is an effective and simple method for affinity maturation of antibodies isolated from a phage antibody library.
Department of Gastroenterology, First Affiliated Hospital, Jinan University, Guangzhou 510630, China.
Xiao-hua
XH
Lu
Dong-hua
DH
Yang
Min
M
Zhou
Shao-hui
SH
Tang
D000912:Antibodies, Neoplasm / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D006528:Carcinoma, Hepatocellular / Q000276:immunology* / Q000473:pathology; D006801:Humans; D007128:Immunoglobulin Fragments / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology*; D008113:Liver Neoplasms / Q000276:immunology* / Q000473:pathology; D009154:Mutation; D019151:Peptide Library
D000912:Antibodies, Neoplasm; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D019151:Peptide Library