Patient-derived phage antibody display library as a source of human recombinant antibodies to candidal hsp 90

Abstract

Human recombinant antibodies to Candida albicans hsp 90 were prepared from the cDNA of the peripheral blood lymphocytes of a patient recently recovered from invasive candidiasis. The phage antibody display library was enriched for high affinity binders against the synthetic peptide NKILKVIRKNIVKK (B). This peptide is a dominant conserved epitope, previously identified as being the most commonly recognized by patients recovered from systemic candidiasis and having antibody to the 47 kDa subcomponent of Candida hsp 90. It is in a central position in the proposed protein binding site of human hsp 90. Successful panning against this peptide was demonstrated by DNA fingerprinting individual clones and showing the emergence of four predominant types. These were examined, in vitro, for activity against both hsp 90 and the specific epitope NKILKVIRKNIVKK by immunoblotting and competitive enzyme-linked immunosorbent assay (ELISA) respectively.