2017 International Conference on
BioNano Innovation 2017 ICBNI

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Industrial Biotechnology Stream

Using phage display to isolate antibodies against membrane proteins

25 September 2017 4:00pm
 

Jones ML*, Alfaleh MA, Kumble S, Yeh M, Howard CB, Mahler SM

 

ARC Training Centre for Biopharmaceutical Innovation
National Biologics Facility
Australian Institute for Bioengineering and Nanotechnology
The University of Queensland
St Lucia, QLD, Australia

 

Monoclonal antibodies (mAbs) currently dominate the pharmaceutical market for biologic drugs; five of the top ten drugs in 2016 were antibodies1 and there are over 250 mAbs in Phase II and III clinical trials.2  The majority of these mAbs target membrane protein biomarkers, an attractive target as the expression levels of these proteins can define certain cell types and disease states.  Therapeutically relevant, human-derived antibodies can be isolated from highly diverse libraries using phage display by exposing the library to the antigen of interest, in a process called biopanning.  Antigen quality is a crucial factor for successful biopanning, as therapeutic antibodies must be able to recognise the native, conformational structure of a protein.  This presentation will provide an overview of different strategies to present antigens to the phage library, including a novel method for panning on whole cells to ensure correct conformational display of membrane proteins.3  This method uses transiently-expressed GFP-tagged membrane proteins, coupled with fluorescence-activated cell sorting (FACS), to ensure a high receptor density and incorporates methods to reduce the enrichment of binders to irrelevant antigens.  Results using cell-surface canine c-Kit protein will be presented.

Flow cytometry analysis of phage pools after one (black), two (red) and three (blue) rounds of biopanning, on C-Kit-GFP transfected cells

References:

  1. http://www.genengnews.com/the-lists/the-top-15-best-selling-drugs-of-2016/77900868
  2. Reichert JM. Antibodies to watch in 2017. MAbs 2017; 9:167.
  3. Jones ML, et al. Targeting membrane proteins for antibody discovery using phage display. Sci Rep 2016; 6:26240

 

Biographic Details

Dr Martina Jones

Title:    Deputy Director, ARC Training Centre for Biopharmaceutical Innovation

            Operations Manager, National Biologics Facility

AIBN, University of Queensland, Australia

Phone: +61 7 3346 3178   E-mail: martina.jones@uq.edu.au

Research interests: antibodies, biologics, cancer biomarkers

 

 

Venue

Room: 
Hawken N201