PT  - JOURNAL ARTICLE
AU  - Jinsung Noh
AU  - Okju Kim
AU  - Yushin Jung
AU  - Haejun Han
AU  - Jung-Eun Kim
AU  - Soohyun Kim
AU  - Sanghyub Lee
AU  - Jaeseong Park
AU  - Rae Hyuck Jung
AU  - Sang il Kim
AU  - Jaejun Park
AU  - Jerome Han
AU  - Hyunho Lee
AU  - Duck Kyun Yoo
AU  - Amos C. Lee
AU  - Euijin Kwon
AU  - Taehoon Ryu
AU  - Junho Chung
AU  - Sunghoon Kwon
TI  - High-throughput retrieval of physical DNA for NGS-identifiable clones in phage display library
AID  - 10.1101/370809
DP  - 2018 Jan 01
TA  - bioRxiv
PG  - 370809
4099  - http://biorxiv.org/content/early/2018/07/18/370809.short
4100  - http://biorxiv.org/content/early/2018/07/18/370809.full
AB  - 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.