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Optimizing Production of Antigens and Fabs in the Context of Generating Recombinant Antibodies to Human Proteins.

Authors :
Nan Zhong
Peter Loppnau
Alma Seitova
Mani Ravichandran
Maria Fenner
Harshika Jain
Anandi Bhattacharya
Ashley Hutchinson
Marcin Paduch
Vincent Lu
Michal Olszewski
Anthony A Kossiakoff
Evan Dowdell
Akiko Koide
Shohei Koide
Haiming Huang
Vincent Nadeem
Sachdev S Sidhu
Jack F Greenblatt
Edyta Marcon
Cheryl H Arrowsmith
Aled M Edwards
Susanne Gräslund
Source :
PLoS ONE, Vol 10, Iss 10, p e0139695 (2015)
Publication Year :
2015
Publisher :
Public Library of Science (PLoS), 2015.

Abstract

We developed and optimized a high-throughput project workflow to generate renewable recombinant antibodies to human proteins involved in epigenetic signalling. Three different strategies to produce phage display compatible protein antigens in bacterial systems were compared, and we found that in vivo biotinylation through the use of an Avi tag was the most productive method. Phage display selections were performed on 265 in vivo biotinylated antigen domains. High-affinity Fabs (

Subjects

Subjects :
Medicine
Science

Details

Language :
English
ISSN :
19326203
Volume :
10
Issue :
10
Database :
Directory of Open Access Journals
Journal :
PLoS ONE
Publication Type :
Academic Journal
Accession number :
edsdoj.18dddf47c540eb9b460223ee62eb80
Document Type :
article
Full Text :
https://doi.org/10.1371/journal.pone.0139695