Affiliations 

  • 1 Institute for Research in Molecular Medicine, University Sains Malaysia, 11800 Penang, Malaysia
  • 2 Institute for Research in Molecular Medicine, University Sains Malaysia, 11800 Penang, Malaysia. Electronic address: [email protected]
Anal Biochem, 2015 May 15;477:56-61.
PMID: 25769419 DOI: 10.1016/j.ab.2015.02.026

Abstract

The low presentation efficiency of Fab (fragment antigen binding) fragments during phage display is largely due to the complexity of disulphide bond formation. This can result in the presentation of Fab fragments devoid of a light chain during phage display. Here we propose the use of a coplasmid system encoding several molecular chaperones (DsbA, DsbC, FkpA, and SurA) to improve Fab packaging. A comparison was done using the Fab fragment from IgG and IgD. We found that the use of the coplasmid during phage packaging was able to improve the presentation efficiency of the Fab fragment on phage surfaces. A modified version of panning using the coplasmid system was evaluated and was successful at enriching Fab binders. Therefore, the coplasmid system would be an attractive alternative for improved Fab presentation for phage display.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.