Integrating SpyCatcher/SpyTag covalent fusion technology into phage display workflows for rapid antibody discovery.

Details

Serval ID
serval:BIB_FCB67A02E15F
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Integrating SpyCatcher/SpyTag covalent fusion technology into phage display workflows for rapid antibody discovery.
Journal
Scientific reports
Author(s)
Fierle J.K., Abram-Saliba J., Brioschi M., deTiani M., Coukos G., Dunn S.M.
ISSN
2045-2322 (Electronic)
ISSN-L
2045-2322
Publication state
Published
Issued date
06/09/2019
Peer-reviewed
Oui
Volume
9
Number
1
Pages
12815
Language
english
Notes
Publication types: Journal Article
Publication Status: epublish
Abstract
An early bottleneck in the rapid isolation of new antibody fragment binders using in vitro library approaches is the inertia encountered in acquiring and preparing soluble antigen fragments. In this report, we describe a simple, yet powerful strategy that exploits the properties of the SpyCatcher/SpyTag (SpyC/SpyT) covalent interaction to improve substantially the speed and efficiency in obtaining functional antibody clones of interest. We demonstrate that SpyC has broad utility as a protein-fusion tag partner in a eukaryotic expression/secretion context, retaining its functionality and permitting the direct, selective capture and immobilization of soluble antigen fusions using solid phase media coated with a synthetic modified SpyT peptide reagent. In addition, we show that the expressed SpyC-antigen format is highly compatible with downstream antibody phage display selection and screening procedures, requiring minimal post-expression handling with no sample modifications. To illustrate the potential of the approach, we have isolated several fully human germline scFvs that selectively recognize therapeutically relevant native cell surface tumor antigens in various in vitro cell-based assay contexts.
Pubmed
Web of science
Open Access
Yes
Create date
17/09/2019 18:06
Last modification date
07/04/2020 6:26
Usage data