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

• Main Authors: Mohamed A. Alfaleh, Martina L. Jones, Christopher B. Howard, Stephen M. Mahler
• Format: Article
• Language: English
• Published: MDPI AG 2017-08-01
• Series: Antibodies
• Subjects: affinity selection; antibodies; cell markers; cell receptors; competitive elution; epitopes; panning; phage display; transfection; whole cell
• Online Access: https://www.mdpi.com/2073-4468/6/3/10

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.