High-throughput retrieval of physical DNA for NGS-identifiable clones in phage display library. Issue 3 (3rd April 2019)

• Record Type: Journal Article
• Authors: Noh, Jinsung; Kim, Okju; Jung, Yushin; Han, Haejun; Kim, Jung-Eun; Kim, Soohyun; Lee, Sanghyub; Park, Jaeseong; Jung, Rae Hyuck; Kim, Sang Il; Park, Jaejun; Han, Jerome; Lee, Hyunho; Yoo, Duck Kyun; Lee, Amos C.; Kwon, Euijin; Ryu, Taehoon; Chung, Junho; Kwon, Sunghoon
• Is Part Of: MAbs. Volume 11:Issue 3(2019)
• Page Start: 532
• Page End: 545
• Publication Date: 2019-04-03
• Subjects: Antibody discovery -- monoclonal antibody -- lead antibody -- antibody library -- phage display -- antibody library sequencing -- NGS -- clone retrieval -- rare clones; Monoclonal antibodies -- Therapeutic use -- Periodicals; Monoclonal antibodies -- Periodicals; Antibodies, Monoclonal -- Periodicals; 616.0798
• DOI: 10.1080/19420862.2019.1571878
• Languages: English
• ISSNs: 1942-0862

Abstract

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 technical limitations, such as low process throughput from the 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™. This new platform 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 library targeting human hepatocyte growth factor protein.