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

2019; Landes Bioscience; Volume: 11; Issue: 3 Linguagem: Inglês

10.1080/19420862.2019.1571878

1942-0870

Jinsung Noh, Okju Kim, Yushin Jung, Haejun Han, Jung‐Eun Kim, Soohyun Kim, Sanghyub Lee, Jaeseong Park, Rae Hyuck Jung, Sang il Kim, Jae-Jun Park, Jerome Han, Hyunho Lee, Duck Kyun Yoo, Amos Chungwon Lee, Euijin Kwon, Taehoon Ryu, Junho Chung, Sunghoon Kwon,

T-cell and B-cell Immunology

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.