Affinity maturation of a single-chain antibody against hepatocellular carcinoma / 中华肝脏病杂志

ABSTRACT

<p><b>OBJECTIVES</b>To obtain a single-chain antibody with high affinity against hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>Error-prone PCR is an effective and simple method for affinity maturation of antibodies isolated from a phage antibody library.</p>