Human monoclonal antibodies to CEA from combinatorial libraries on phage surfaces

Objective To construct human phage antibody library and to produce human monoclonal antibodies to CEA. Methods Human phage antibody library was obtained from lymphonode of ten patients with colon carcinoma. Concentrations of plasma CEA of each patient was more than 10 ng/ml. Employing routing RT-PCR with sets of oligo nucleotide primers designed by Kang, the genes of human immunoglobulin Fd and k light chains were amplified and cloned into phagemids (p3MH) to generate human phage antibody library. Human CEA as the coated antigen was used to pan the phage antibody library. Results A phage antibody library ofκchain containing 1.4×107 members was constructed and a phage antibody library of Fab had 5.2×106 members. The VH gene segment was derived from VH3 gene family and Vκgene segment from the Vκgene family by Internet GenBank comparison. Conclusion The success of isolating human anti-CEA Fab prove the usefulness of phage display system in human antibody preparation.