New avenues for phage-display library to produce a Cryptococcus-specific anti-idiotypic antibody of HM-1 killer toxin.

摘要: Existing antifungal drugs are notable for their inability to act rapidly, as well toxicity and limited spectrum. The identification of fungal-specific genes virulence factors would provide targets new influential drugs. display repertories antibody fragments on the surface filamentous phage offers a way produce immunoreagents defined specificities. Here we report selection Cryptococcus-specific by using phage-display panning from cDNA library, where bactericidal antibodies have been developed against conserved surface-exposed antigens. A single-chain variable fragment (scFv) library was constructed splenocyte an immunized mouse idiotypic vaccination with HM-1 killer toxin (HM-1) neutralizing monoclonal (nmAb-KT) that used Cryptococcus neoformans membrane fraction (CnMF). Key elements were antigen (nmAb-KT CnMF) release bound phages competitive elution CnMF at neutral pH condition. Isolated scFvs react specifically C. some other pathogenic non-pathogenic fungal strain's cell wall receptors exerting strong activity in vitro. high affinity clone, designated M1 selected detailed characterization tested anti-cryptococcal IC50 values 5.33 × 10−7 5.56 × 10−7 M neoformans. method described here is technique isolation specific immunoreactive form scFv contained associated proteins. Copyright © 2011 John Wiley & Sons, Ltd.