Selection and Purification of Human Phage-Antibody (Phab) Against HBsAg

Aim: To obtain Fab fragment of human antibody against HBsAg. Methods: Gene of Phab against HBsAg was selected from constructed phage library of human antibodies by using HBsAg-coated ELISA plate and transfected into E. coli to express. Then, soluble Fab fragment was purified by using Actisep Elution Medium as eluant to elute split E. coli through the HiTrap column coupled with goat antibody against human Fab. The purity and the biological activity of the purified Fab fragment were determined respectively by SDA-PAGE gel electrophoresis of protein (Western Blot) and RIA. The combining ability of the Fab fragment with HBsAg was tested by ELISA method. Results: The expression of the Fab fragment against HBsAg by transfected E. coli was confimed by Western Blot and ELISA; the purity of the Fab fragment reach the electrophoretically pure, and the product had quite high activity. Conclusion: The selection and purification of human antibody against HBsAg from phage--antibody library lay a sound foundation of further studying.