Construction of human immunoglobulin combinatorial library and screening of phage antibodies to hepatitis A virus

Objective To construct human phage antibody library and produce human monoclonal antibodies. Methods Human immunoglobulin genes were amplified by RT PCR and cloned into phage display system (pComb3/VCSM13) to generate phage antibody library. Antibodies against hepatitis A virus were screened by biopanning. Results A human combinatorial antibody library consisting of 6 93×10 7 independent clones was generated with a titer of 8×10 14 PFU/ml. Forty percent of the clones contained Fab genes. Three rounds of selection against McAb captured HAV particles showed specific enrichment of phage antibodies. Direct ELISA and competitive inhibition ELISA verified that the positive phage antibodies were highly specific against HAV. Conclusion The success of isolating human anti HAV Fab prove the usefulness of phage display system in human McAb preparation.