Improvement of affinity of phage engineered antibody against HBsAg through shuffling light chain gene library / 解放军医学杂志

ABSTRACT

Objective To explore the effect of light chain gene library slauffling on the affinity of phage engineered antibody against HBsAg. Methods Human antibody light chain gene repertoire generated by RT-PCR from human peripheral blood lymphocyte, was inserted into the phagmid containing Fd gene to construct the phage antibody sublibrary, from which the light chain gene matching the heavy chain Fd gene was screened. Results After three rounds of selection by biopanning, eight clones with higher absorbency than that of original clone at 490nm in ELISA were obtained, indicating that the affinity of chain shuffled phage antibodies (phab) was improved (A_ 490 from 0.43?0.09 to 1.24?0.10). DNA sequencing showed three of the five V_L genes were ? type, and the other two were ? type. Conclusions The phab obtained possesses the specificity of anti-HBsAg property.