title: Targeting membrane proteins for antibody discovery using phage display.
abstract: A critical factor in the successful isolation of new antibodies by phage display is the presentation of a correctly folded antigen. While this is relatively simple for soluble proteins which can be purified and immobilized onto a plastic surface, membrane proteins offer significant challenges for antibody discovery. Whole cell panning allows presentation of the membrane protein in its native conformation, but is complicated by a low target antigen density, high background of irrelevant antigens and non-specific binding of phage particles to cell surfaces. The method described here uses transient transfection of alternating host cell lines and stringent washing steps to address each of these limitations. The successful isolation of antibodies from a naive scFv library is described for three membrane bound proteins; human CD83, canine CD117 and bat CD11b.
authors.affiliation: Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia Queensland 4072 Australia.
authors.forename: Martina L
authors.initials: ML
authors.lastname: Jones
authors.affiliation: Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia Queensland 4072 Australia.|Faculty of Pharmacy; King Abdulaziz University, 21589 Jeddah, Saudi Arabia.
authors.forename: Mohamed A
authors.initials: MA
authors.lastname: Alfaleh
authors.affiliation: Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia Queensland 4072 Australia.
authors.forename: Sumukh
authors.initials: S
authors.lastname: Kumble
authors.affiliation: Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia Queensland 4072 Australia.
authors.forename: Shuo
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia Queensland 4072 Australia.|Queensland Brain Institute, The University of Queensland, St Lucia Queensland 4072 Australia.
authors.forename: Geoffrey W
authors.initials: GW
authors.lastname: Osborne
authors.affiliation: Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia Queensland 4072 Australia.
authors.forename: Michael
authors.initials: M
authors.lastname: Yeh
authors.affiliation: Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia Queensland 4072 Australia.
authors.forename: Neetika
authors.initials: N
authors.lastname: Arora
authors.affiliation: Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia Queensland 4072 Australia.
authors.forename: Jeff Jia Cheng
authors.initials: JJ
authors.lastname: Hou
authors.affiliation: Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia Queensland 4072 Australia.
authors.forename: Christopher B
authors.initials: CB
authors.lastname: Howard
authors.affiliation: Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia Queensland 4072 Australia.
authors.forename: David Y
authors.initials: DY
authors.lastname: Chin
authors.affiliation: Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia Queensland 4072 Australia.
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Mahler
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D015703:Antigens, CD / Q000276:immunology; D039481:CD11b Antigen / Q000276:immunology; D016466:CHO Cells; D061505:Cell Surface Display Techniques / Q000379:methods*; D002685:Chiroptera; D003412:Cricetulus; D004285:Dogs; D057809:HEK293 Cells; D006801:Humans; D007136:Immunoglobulins / Q000276:immunology; D008562:Membrane Glycoproteins / Q000276:immunology; D008565:Membrane Proteins / Q000276:immunology*; D019151:Peptide Library; D019009:Proto-Oncogene Proteins c-kit / Q000276:immunology; D014162:Transfection; D000097802:CD83 Antigen
chemical_list: D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D039481:CD11b Antigen; D007136:Immunoglobulins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D019151:Peptide Library; D019009:Proto-Oncogene Proteins c-kit; D000097802:CD83 Antigen