PHAGE DISPLAY USED FOR GENE CLONING OF HUMAN RECOMBINANT ANTIBODY AGAINST THE ERYTHROCYTE SURFACE-ANTIGEN, RHESUS-D

A novel phage display system has been developed for PCR amplification and cloning of the Fab fragments of human immunoglobulin genes. Using this system, we have cloned an antibody from a mouse-human hybridoma c ell line directed against the erythrocyte antigen rhesus D. Intact ery throcytes were used for absorption of the Fab phages. Soluble Fab frag ments produced from the cloned material showed identical performance t o the parental antibody in agglutination assays. Gel filtration confir med that the Fab fragment consists of a kappa-Fd heterodimer. The succ essful use of intact cells for selection of specific Fab phages demons trates that it is possible to by-pass purification of the antigen of i nterest. Comparison with published germline sequences demonstrated tha t the immunoglobulin coding regions had the highest homology to the V- H 1.9III and V, Hum kappa v325 germline genes, respectively.