Adapter-directed display: a modular design for shuttling display on phage surfaces.
A novel adapter-directed phage display system was developed with modular features. In this system, the target protein is expressed as a fusion protein consisting of adapter GR1 from the phagemid vector, while the recombinant phage coat protein is expressed as a fusion protein consisting of adapter GR2 in the helper phage vector. Surface display of the target protein is accomplished through specific heterodimerization of GR1 and GR2 adapters, followed by incorporation of the heterodimers into phage particles. A series of engineered helper phages were constructed to facilitate both display valency and formats, based on various phage coat proteins. As the target protein is independent of a specific phage coat protein, this modular system allows the target protein to be displayed on any given phage coat protein and allows various display formats from the same vector without the need for reengineering. Here, we demonstrate the shuttling display of a single-chain Fv antibody on phage surfaces between multivalent and monovalent formats, as well as the shuttling display of an antigen-binding fragment molecule on phage coat proteins pIII, pVII, and pVIII using the same phagemid vectors combined with different helper phage vectors. This adapter-directed display concept has been applied to eukaryotic yeast surface display and to a novel cross-species display that can shuttle between prokaryotic phage and eukaryotic yeast systems.
Wang
Kevin Caili
KC
Abmaxis Inc., a wholly owned subsidiary of Merck & Co, Inc, WP26-413, 770 Sumneytown Pike, West Point, PA 19486, USA. kevin_wang@merck.com
Wang
Xinwei
X
Zhong
Pingyu
P
Luo
Peter Peizhi
PP
D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000235:genetics; D001483:Base Sequence; D036022:Capsid Proteins / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D005818:Genetic Engineering; D005822:Genetic Vectors; D006378:Helper Viruses / Q000235:genetics; D007140:Immunoglobulin Fab Fragments / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D008969:Molecular Sequence Data; D019151:Peptide Library; D053660:Receptors, Interleukin-13 / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics
D036022:Capsid Proteins; D017931:DNA Primers; D007140:Immunoglobulin Fab Fragments; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D053660:Receptors, Interleukin-13; D011993:Recombinant Fusion Proteins