Multivalent display system on filamentous bacteriophage pVII minor coat protein.
The systems for display of foreign peptides and polypeptides on filamentous bacteriophage have exploited genetic fusion to all of the five coat proteins. Multivalent display systems allowing selection of low affinity antibody fragments have been devised for fusions to gene III. However, since pIII has to interact with the bacterial receptors during the infection process, reduced infectivity can be observed. Alternative display systems utilizing other coat protein have been examined. These, however, take advantage of phagemid systems, in which a mixture of fusion and non-fusion coat proteins becomes displayed, thus preventing multivalent display. In this paper, we describe genetically stable fusion of scFv fragments to gene VII directly in the phage genome, thus giving rise to a multivalent display system where infectivity is not comprised. A hundred-fold enrichments factor can be obtained in model selection. Our results demonstrate that the small size of pVII (33 amino acids) is not structurally compromised by fusion of scFv antibody fragments at their N-terminus, thus demonstrating the feasibility of utilizing pVII as a fusion partner.
Kwaśnikowski
Piotr
P
Institute of Bioorganic Chemistry, Poznań, Poland. pkk@ibch.poznan.pl
Kristensen
Peter
P
Markiewicz
Wojciech T
WT
D000818:Animals; D017104:Bacteriophage M13 / Q000235:genetics*; D015153:Blotting, Western; D036022:Capsid Proteins / Q000235:genetics*; D002352:Carrier Proteins / Q000235:genetics / Q000276:immunology; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000821:virology; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics* / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000276:immunology; D019151:Peptide Library; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D012710:Serum Albumin, Bovine / Q000276:immunology; D019809:Streptavidin / Q000378:metabolism; D025801:Ubiquitin / Q000276:immunology
D036022:Capsid Proteins; D002352:Carrier Proteins; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D011993:Recombinant Fusion Proteins; D025801:Ubiquitin; D012710:Serum Albumin, Bovine; D019809:Streptavidin