Affinity maturation of a single-chain antibody against hepatocellular carcinoma.
- Author:
Xiao-hua LU
1
;
Dong-hua YANG
;
Min ZHOU
;
Shao-hui TANG
Author Information
- Publication Type:Journal Article
- MeSH: Antibodies, Neoplasm; immunology; Antibody Affinity; Antibody Specificity; Carcinoma, Hepatocellular; immunology; pathology; Humans; Immunoglobulin Fragments; immunology; Immunoglobulin Variable Region; genetics; immunology; Liver Neoplasms; immunology; pathology; Mutation; Peptide Library
- From: Chinese Journal of Hepatology 2006;14(3):192-195
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVESTo obtain a single-chain antibody with high affinity against hepatocellular carcinoma (HCC).
METHODSA 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.
RESULTSThe 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.
CONCLUSIONError-prone PCR is an effective and simple method for affinity maturation of antibodies isolated from a phage antibody library.
