EFFICIENT DISPLAY OF AN HCV CDNA EXPRESSION LIBRARY AS C-TERMINAL FUSION TO THE CAPSID PROTEIN-D OF BACTERIOPHAGE-LAMBDA

We describe the construction and characterization of a hepatitis C vir us (HCV) cDNA expression library displayed as a fusion to the carboxy terminus of the capsid protein D of bacteriophage lambda, cDNA inserts were obtained by tagged random-priming of the HCV genome and cloned i nto a lambda vector from which chimeric phage bearing both wild-type D protein and D fusion products on the capsid surface were produced. Th e resulting library was affinity-selected with anti-HCV human monoclon al antibodies recognizing linear or conformational epitopes, and human sera from HCV-infected patients. Selection was monitored by immune-sc reening experiments, ELISA, and sequence analysis of positive clones. The performance of this Library was compared with two additional HCV c DNA display libraries generated as N-terminal fusions to the III and W I capsid proteins of filamentous phage M13. The results obtained demon strate the great potential of the lambda display system for constructi ng complex cDNA libraries for natural ligand discovery. (C) 1998 Acade mic Press.