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高通量检索噬菌体展示文库中可用于 NGS 鉴定克隆的物理 DNA。

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

机构信息

a Department of Electrical Engineering and Computer Science , Seoul National University , Seoul , Republic of Korea.

b Bioengineering Research Institute, Celemics, Inc , Seoul , Republic of Korea.

出版信息

MAbs. 2019 Apr;11(3):532-545. doi: 10.1080/19420862.2019.1571878. Epub 2019 Feb 12.

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.

摘要

在抗体发现中,深入分析抗体文库和高通量检索文库中的克隆对于识别和利用具有不同特性的稀有克隆至关重要。然而,现有的方法存在技术局限性,例如克隆过程繁琐导致的低过程通量,以及对显性克隆进行不必要的重复测试导致的表型筛选能力浪费。为了克服这些局限性,我们开发了一种新的高通量文库克隆识别和检索平台,TrueRepertoireTM。这个新平台提供了高度准确的克隆序列,以及抗体片段重链和轻链之间的连接信息。此外,还可以根据序列信息以高通量方式检索克隆的物理 DNA。我们验证了序列的高度准确性,并证明没有平台特异性偏差。此外,我们还通过针对人肝细胞生长因子蛋白的噬菌体展示单链可变片段文库验证了 TrueRepertoireTM 的适用性。

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