Targeting membrane proteins for antibody discovery using phage display

Jones, Martina L., Alfaleh, Mohamed A., Kumble, Sumukh, Zhang, Shuo, Osborne, Geoffrey W., Yeh, Michael, Arora, Neetika, Hou, Jeff Jia Cheng, Howard, Christopher B., Chin, David Y. and Mahler, Stephen M. (2016) Targeting membrane proteins for antibody discovery using phage display. Scientific Reports, 6 . doi:10.1038/srep26240

Author Jones, Martina L.
Alfaleh, Mohamed A.
Kumble, Sumukh
Zhang, Shuo
Osborne, Geoffrey W.
Yeh, Michael
Arora, Neetika
Hou, Jeff Jia Cheng
Howard, Christopher B.
Chin, David Y.
Mahler, Stephen M.
Title Targeting membrane proteins for antibody discovery using phage display
Journal name Scientific Reports   Check publisher's open access policy
ISSN 2045-2322
Publication date 2016-05-18
Sub-type Article (original research)
DOI 10.1038/srep26240
Open Access Status DOI
Volume 6
Total pages 11
Place of publication London, United Kingdom
Publisher Nature Publishing Group
Collection year 2017
Language eng
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.
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ
Additional Notes Article number 26240

Document type: Journal Article
Sub-type: Article (original research)
Collections: HERDC Pre-Audit
Queensland Brain Institute Publications
Australian Institute for Bioengineering and Nanotechnology Publications
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