Affinity maturation of a single-chain antibody for hepatocellular carcinoma by error-prone PCR and DNA shuffling
- VernacularTitle:错配PCR和DNA改组技术提高抗肝癌单链抗体亲和力
- Author:
Xiaohua LU
;
Donghua YANG
;
Shaohui TANG
;
Min ZHOU
- Publication Type:Journal Article
- Keywords:
Error-prone PCR;
DNA shuffling;
Liver neoplasms;
Antibody affinity
- From:
Chinese Journal of Pathophysiology
1989;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To obtain a single-chain antibody with high affinity for hepatocellular carcinoma (HCC). METHODS: A second single-chain antibody mutant library was established by using error-prone PCR and DNA shuffling. Single-chain antibodies with high affinity for hepatocellular carcinoma were selected from phage antibody library by using ELISA. RESULTS: The content of the second single-chain antibody mutant library was about 4.5?10~7. Two selected mutants M25 and M36 were obtained after 3 rounds of panning and ELISA. Immunoassay showed that M25 and M36 bound to human HCC cells specifically. The relative affinity of M25 was 2.0 folds higher than that of the original antibody, and M36 was 2.4 folds higher than the original antibody. CONCLUSION: Error-prone PCR combined with DNA shuffling is an effective method to improve affinity of antibodies isolated from phage antibody library.
