Application of a Synthetic Phage Antibody Library (ETH-2) for the Isolation of Single Chain Fragment Variable (scFv) Human Antibodies to the Pathogenic Isoform of the Hamster Prion Protein (HaPrPsc)

Abstract

To overcome the limitation represented by the poor immunogenicity of prion protein (PrP) for conventional monoclonal antibodies preparation, we adopted an antibody phage display strategy to isolate specific human single chain fragment variable (scFv) directed towards the pathogenic isoform of the hamster prion protein (HaPrPsc). Phage-displaying HaPrPsc reactive scFvs were obtained after three rounds of selection of the ETH- 2 synthetic antibody library on HaPrPsc-coated immunotubes and subsequent amplification in TG1 E. coli cells. These phage-antibodies bind in ELISA to HaPrPsc and do not cross-react with the recombinant hamster prion protein (rHaPrP). Sequence analyses of the gene encoding for the antibody fragments and antigen recognition patterns determined by flow-cytometry on lymphoid cells indicate that the selected scFv recognize distinct epitopes in the PrPsc molecule. The results of this study demonstrate that display of scFvs on filamentous phage offers the possibility of producing phage antibodies showing immunoglobulin-like functions using only in vitro procedures, thus overcoming limitations of conventional hybridoma technology.