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High-throughput retrieval of physical DNA for NGS-identifiable clones in phage display library

Authors :
Euijin Kwon
Hyunho Lee
Junho Chung
Sang Hyub, Lee
Yushin Jung
Jerome Han
Jaeseong Park
Rae Hyuck Jung
Jinsung Noh
Sunghoon Kwon
Duck Kyun Yoo
Amos Chungwon Lee
Haejun Han
Soohyun Kim
Jung-Eun Kim
Okju Kim
Jaejun Park
Taehoon Ryu
Sang Il Kim
Source :
mAbs. 11:532-545
Publication Year :
2019
Publisher :
Informa UK Limited, 2019.

Abstract

In antibody discovery, in-depth analysis of an antibody library and high-throughput retrieval of clones in the library are crucial to identifying and exploiting rare clones with different properties. However, existing methods have several technical limitations such as low process throughput from laborious cloning process and waste of the phenotypic screening capacity from unnecessary repetitive tests on the dominant clones. To overcome the limitations, we developed a new high-throughput platform for the identification and retrieval of clones in the library, TrueRepertoire™. TrueRepertoire™ provides highly accurate sequences of the clones with linkage information between heavy and light chains of the antibody fragment. Additionally, the physical DNA of clones can be retrieved in high throughput based on the sequence information. We validated the high accuracy of the sequences and demonstrated that there is no platform-specific bias. Moreover, the applicability of TrueRepertoire™ was demonstrated by a phage-displayed single-chain variable fragment (scFv) library targeting human hepatocyte growth factor (hHGF) protein.

Details

ISSN :
19420870 and 19420862
Volume :
11
Database :
OpenAIRE
Journal :
mAbs
Accession number :
edsair.doi.dedup.....534be3ba5e75265b5bbebdc8861db6ef
Full Text :
https://doi.org/10.1080/19420862.2019.1571878