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[Affinity maturation of a single-chain antibody against hepatocellular carcinoma]
Xiao-hua Lu1, Dong-hua Yang, Min Zhou
1Department of Gastroenterology, First Affiliated Hospital, Jinan University, Guangzhou 510630, China.
Objectives:
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.
Conclusion:
Error-prone PCR is an effective and simple method for affinity maturation of antibodies isolated from a phage antibody library.
