1. High-throughput retrieval of physical DNA for NGS-identifiable clones in phage display library
- Author
-
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, and Sang Il Kim
- Subjects
Phage display ,Computer science ,medicine.drug_class ,Phenotypic screening ,Immunology ,Computational biology ,Immunoglobulin light chain ,Monoclonal antibody ,Avian Proteins ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Report ,medicine ,Animals ,Immunology and Allergy ,Bacteriophages ,Throughput (business) ,030304 developmental biology ,Cloning ,0303 health sciences ,Human hepatocyte ,chemistry ,030220 oncology & carcinogenesis ,Cell Surface Display Techniques ,Chickens ,DNA ,Single-Chain Antibodies - 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.
- Published
- 2019
- Full Text
- View/download PDF