Improved Fab presentation on phage surface with the use of molecular chaperone coplasmid system

Loh, Qiuting; Leong, Siew Wen; Tye, Gee Jun; Choong, Yee Siew; Lim, Theam Soon

doi:10.1016/j.ab.2015.02.026

Cited by 7 publications

(3 citation statements)

References 24 publications

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“…Fabs are often reported as having relatively low phage display levels compared to scFvs, which may be attributed to their relatively complex disulphide bond formation [26, 34]. Several recent studies have looked to improve Fab display levels.…”

Section: Antigen Binding Fragment (Fab)mentioning

confidence: 99%

“…For example, co-plasmid expression of DsbA , DsbC , FkpA and SurA has been used. The expression of the peptidyl prolyl cis–trans isomerases (PPIases) FkpA and SurA directed to the periplasm may improve folding, and DsbA and DsbC help optimise disulphide bond formation [34]. The selection of Fabs during panning was greatly improved with much higher levels of Fab detected on phage through each round of panning compared to when the co-plasmid was not present.…”

Section: Antigen Binding Fragment (Fab)mentioning

confidence: 99%

“…Both advances will facilitate the optimal cloning of diversity into libraries in the future. The co-expression of chaperones and isomerases [34, 36] as well as the use of the co-translation secretion pathway with optimised signal sequences [25] can significantly improve antibody phage display efficiency, thereby improving the efficiency of isolating binders. These same strategies also improve soluble antibody expression levels.…”

Section: Conclusion and Future Perspectivesmentioning

confidence: 99%

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Advances in the Production and Batch Reformatting of Phage Antibody Libraries

et al. 2019

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Phage display antibody libraries have proven an invaluable resource for the isolation of diagnostic and potentially therapeutic antibodies, the latter usually being antibody fragments converted into IgG formats. Recent advances in the production of highly diverse and functional antibody libraries are considered here, including for Fabs, scFvs and nanobodies. These advances include codon optimisation during generation of CDR diversity, improved display levels using novel signal sequences, molecular chaperones and isomerases and the use of highly stable scaffolds with relatively high expression levels. In addition, novel strategies for the batch reformatting of scFv and Fab phagemid libraries, derived from phage panning, into IgG formats are described. These strategies allow the screening of antibodies in the end-use format, facilitating more efficient selection of potential therapeutics.

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Section: Antigen Binding Fragment (Fab)mentioning

confidence: 99%

Section: Antigen Binding Fragment (Fab)mentioning

confidence: 99%