4.7 Article

A High-Throughput Single-Clone Phage Fluorescence Microwell Immunoassay and Laser-Driven Clonal Retrieval System

期刊

BIOMOLECULES
卷 10, 期 4, 页码 -

出版社

MDPI
DOI: 10.3390/biom10040517

关键词

phage display; single clone; microchip

资金

  1. National Research Foundation of Korea (NRF) [2017R1A6A3A04010692, 2016M3A9B5941738]
  2. Korea Drug Development Fund [KDDF-201904-19]
  3. Global Research Development Center Program through the NRF - Ministry of Science and ICT (MSIT) [2015K1A4A3047345]
  4. Korea Evaluation Institute of Industrial Technology (KEIT) [KDDF-201904-19] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
  5. National Research Foundation of Korea [2017R1A6A3A04010692, 2016M3A9B5941738] 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

Phage display is one of the most frequently used platform technologies utilized to screen and select therapeutic antibodies, and has contributed to the development of more than 10 therapeutic antibodies used in the clinic. Despite advantages like efficiency and low cost, it has intrinsic technical limitations, such as the asymmetrical amplification of the library after each round of biopanning, which is regarded as a reason for it yielding a very limited number of antigen binders. In this study, we developed a high-throughput single-clonal screening system comprised of fluorescence immunoassays and a laser-driven clonal DNA retrieval system using microchip technology. Using this system, from a single-chain variable fragment (scFv) library displayed on phages with a complexity of 5.21 x 10(5) harboring random mutations at five amino acid residues, more than 70,000 clones-corresponding to similar to 14% of the library complexity-were screened, resulting in 78 antigen-reactive scFv sequences with mutations restricted to the randomized residues. Our results demonstrate that this system can significantly reduce the number of biopanning rounds, or even eliminate the need for this process for libraries with lower complexity, providing an opportunity to obtain more diverse clones from the library.

作者

我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。

评论

主要评分

4.7
评分不足

次要评分

新颖性
-
重要性
-
科学严谨性
-
评价这篇论文

推荐

暂无数据
暂无数据