Targeting membrane proteins for antibody discovery using phage display

• DOI: 10.1038/srep26240
• Publication Date: 2016-05-18T09:32:49Z
• Authors: Martina L. Jones; Mohamed A. Alfaleh; Sumukh Kumble; Shuo Zhang; Geoffrey W. Osborne; Michael Yeh; Neetika Arora; Jeff Jia Cheng Hou; Christopher B. Ho...; David Y. Chin; Stephen M. Mahler
• Keywords: Panning (audio)

Abstract A critical factor in the successful isolation of new antibodies by phage display is presentation a correctly folded antigen. While this relatively simple for soluble proteins which can be purified and immobilized onto plastic surface, membrane offer significant challenges antibody discovery. Whole cell panning allows protein its native conformation, but complicated low target antigen density, high background irrelevant antigens non-specific binding particles to surfaces. The method described here uses transient transfection alternating host lines stringent washing steps address each these limitations. from naive scFv library three bound proteins; human CD83, canine CD117 bat CD11b.