4.5 Article

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

期刊

MABS
卷 11, 期 3, 页码 532-545

出版社

TAYLOR & FRANCIS INC
DOI: 10.1080/19420862.2019.1571878

关键词

Antibody discovery; monoclonal antibody; lead antibody; antibody library; phage display; antibody library sequencing; NGS; clone retrieval; rare clones

资金

  1. National Research Foundation of Korea (NRF) - Ministry of Science and ICT(MSIT) [2015K1A4A3047345]
  2. Brain Korea 21 Plus Project in 2018
  3. Korea Health Technology R&D Project of the Korea Health Industry Development Institute (KHIDI) - Ministry of Health & Welfare, Republic of Korea [HI18C2282]
  4. Bio & Medical Technology Development Program of the National Research Foundation (NRF) - Korean government (MSIT) [2018M3A9D7079488]
  5. National Research Foundation of Korea [2018M3A9D7079488, 2015K1A4A3047345] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)

向作者/读者索取更多资源

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 (TM). 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 (TM) was demonstrated by a phage-displayed single-chain variable fragment library targeting human hepatocyte growth factor protein.

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