[Selection of human phage antibodies against HAV and DNA sequence analysis].

ABSTRACT

OBJECTIVE: To obtain the anti-HAV antibodies. METHODS: The antibody-capturing antigen strategy was used in panning antibody library with impure antigen. The HAV-binding activity and specificity of phage antibodies were assayed by sandwich ELISA and competitive inhibition ELISA. RESULTS: Phage antibodies(Fab) against HAV were screened from a human immunoglobulin combinatorial library. Fd and light chain genes were sequenced. Comparison of the Fd gene and light chain gene with KABAT database showed that the heavy chain belonged to IgG1 subclass, and its variable region was derived from rearranged germ-line gene of DP88, D3-3 and JH5; while the light chain was a member of V kappa III family, the germ-line gene was DP kappa 22 and J kappa 4. CONCLUSION: Phage antibodies against HAV were selected from the antibody library by using the antibody-capturing antigen strategy. The result also suggested that the antibody library was constructed successfully.