title: Improved Fab presentation on phage surface with the use of molecular chaperone coplasmid system.
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.
authors.affiliation: Institute for Research in Molecular Medicine, University Sains Malaysia, 11800 Penang, Malaysia.
authors.forename: Qiuting
authors.initials: Q
authors.lastname: Loh
authors.affiliation: Institute for Research in Molecular Medicine, University Sains Malaysia, 11800 Penang, Malaysia.
authors.forename: Siew Wen
authors.initials: SW
authors.lastname: Leong
authors.affiliation: Institute for Research in Molecular Medicine, University Sains Malaysia, 11800 Penang, Malaysia.
authors.forename: Gee Jun
authors.initials: GJ
authors.lastname: Tye
authors.affiliation: Institute for Research in Molecular Medicine, University Sains Malaysia, 11800 Penang, Malaysia.
authors.forename: Yee Siew
authors.initials: YS
authors.lastname: Choong
authors.affiliation: Institute for Research in Molecular Medicine, University Sains Malaysia, 11800 Penang, Malaysia. Electronic address: theamsoon@usm.my.
authors.forename: Theam Soon
authors.initials: TS
authors.lastname: Lim
mesh_terms: D007072:Immunoglobulin D / Q000235:genetics*; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D007074:Immunoglobulin G / Q000235:genetics*; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000378:metabolism*; D019151:Peptide Library; D010957:Plasmids / Q000235:genetics*; D011487:Protein Conformation
chemical_list: D007072:Immunoglobulin D; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D018832:Molecular Chaperones; D019151:Peptide Library
keywords: Antibody phage display; Coplasmid; Fab presentation; Molecular chaperone