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Antibodies 2017, 6(3), 10; doi:10.3390/antib6030010

Strategies for Selecting Membrane Protein-Specific Antibodies using Phage Display with Cell-Based Panning

1
Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Brisbane, Queensland 4072, Australia
2
Faculty of Pharmacy, King Abdulaziz University, Jeddah 21589, Saudi Arabia
3
Australian Research Council Training Centre for Biopharmaceutical Innovation, The University of Queensland, Brisbane, Queensland 4072, Australia
4
Centre for Advanced Imaging, The University of Queensland, Brisbane, Queensland 4072, Australia
*
Authors to whom correspondence should be addressed.
Academic Editor: Dimiter S. Dimitrov
Received: 14 June 2017 / Revised: 5 July 2017 / Accepted: 7 July 2017 / Published: 5 August 2017
View Full-Text   |   Download PDF [2872 KB, uploaded 5 August 2017]   |  

Abstract

Membrane proteins are attractive targets for monoclonal antibody (mAb) discovery and development. Although several approved mAbs against membrane proteins have been isolated from phage antibody libraries, the process is challenging, as it requires the presentation of a correctly folded protein to screen the antibody library. Cell-based panning could represent the optimal method for antibody discovery against membrane proteins, since it allows for presentation in their natural conformation along with the appropriate post-translational modifications. Nevertheless, screening antibodies against a desired antigen, within a selected cell line, may be difficult due to the abundance of irrelevant organic molecules, which can potentially obscure the antigen of interest. This review will provide a comprehensive overview of the different cell-based phage panning strategies, with an emphasis placed on the optimisation of four critical panning conditions: cell surface antigen presentation, non-specific binding events, incubation time, and temperature and recovery of phage binders. View Full-Text
Keywords: affinity selection; antibodies; cell markers; cell receptors; competitive elution; epitopes; panning; phage display; transfection; whole cell affinity selection; antibodies; cell markers; cell receptors; competitive elution; epitopes; panning; phage display; transfection; whole cell
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Alfaleh, M.A.; Jones, M.L.; Howard, C.B.; Mahler, S.M. Strategies for Selecting Membrane Protein-Specific Antibodies using Phage Display with Cell-Based Panning. Antibodies 2017, 6, 10.

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