Streamlined human antibody generation and optimization by exploiting designed immunoglobulin loci in a B cell line.
Streamlined human antibody generation and optimization by exploiting designed immunoglobulin loci in a B cell line.
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DOI：
10.1038/s41423-020-0440-9
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发表时间：
2021-06
影响因子：
24.1
通讯作者：
Nakazaki Y
中科院分区：
文献类型：
--
作者：
Seo H;Masuda H;Asagoshi K;Uchiki T;Kawata S;Sasaki G;Yabuki T;Miyai S;Takahashi N;Hashimoto SI;Sawada A;Takaiwa A;Koyama C;Tamai K;Kurosawa K;Lin KY;Ohta K;Nakazaki Y
单克隆抗体（mAb）被广泛用作各种疾病的治疗药物，例如癌症、自身免疫性疾病和感染性疾病。使用禽源性B细胞系DT 40，我们先前开发了抗体展示技术，即ADLi B系统，其快速产生抗原特异性mAb。在这里，我们报告了ADLib系统的人类版本的开发，并展示了功能性人类mAb的简化生成和优化。定制文库首先通过用设计的人对应物替换内源性免疫球蛋白基因来构建。从这些文库中，可以分离产生针对不同抗原的全长人IgG的克隆，如通过选择拮抗性mAb所例示的。利用鸟类生物学，通过将亲本克隆无缝多样化为二级文库，然后进行单细胞分选，以简单的方式实现有效的亲和力成熟，快速提供具有改善的亲和力和功能的mAb。总的来说，我们证明了人类ADLib系统可以作为一个具有独特多样性的综合平台，用于快速从头生成和优化治疗或诊断抗体先导物。此外，我们的研究结果表明，库可以通过导入外源基因到DT 40细胞，表明ADLib系统有可能被应用于快速和有效的定向进化和优化蛋白质在生物医学以外的各个领域。
Monoclonal antibodies (mAbs) are widely utilized as therapeutic drugs for various diseases, such as cancer, autoimmune diseases, and infectious diseases. Using the avian-derived B cell line DT40, we previously developed an antibody display technology, namely, the ADLib system, which rapidly generates antigen-specific mAbs. Here, we report the development of a human version of the ADLib system and showcase the streamlined generation and optimization of functional human mAbs. Tailored libraries were first constructed by replacing endogenous immunoglobulin genes with designed human counterparts. From these libraries, clones producing full-length human IgGs against distinct antigens can be isolated, as exemplified by the selection of antagonistic mAbs. Taking advantage of avian biology, effective affinity maturation was achieved in a straightforward manner by seamless diversification of the parental clones into secondary libraries followed by single-cell sorting, quickly affording mAbs with improved affinities and functionalities. Collectively, we demonstrate that the human ADLib system could serve as an integrative platform with unique diversity for rapid de novo generation and optimization of therapeutic or diagnostic antibody leads. Furthermore, our results suggest that libraries can be constructed by introducing exogenous genes into DT40 cells, indicating that the ADLib system has the potential to be applied for the rapid and effective directed evolution and optimization of proteins in various fields beyond biomedicine.
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影响因子：
14.9
作者：
Kanayama, N;Todo, K;Ohmori, H
通讯作者：
Ohmori, H
影响因子：
10.5
作者：
MCCORMACK, WT;THOMPSON, CB
通讯作者：
THOMPSON, CB
影响因子：
5.3
作者：
Ching KH;Collarini EJ;Abdiche YN;Bedinger D;Pedersen D;Izquierdo S;Harriman R;Zhu L;Etches RJ;van de Lavoir MC;Harriman WD;Leighton PA
通讯作者：
Leighton PA
影响因子：
14.9
作者：
Langer, SJ;Ghafoori, AP;Leinwand, L
通讯作者：
Leinwand, L
DOI：
10.1073/pnas.1114010108
发表时间：
2011-12-20
影响因子：
11.1
作者：
Bowers, Peter M.;Horlick, Robert A.;King, David J.
通讯作者：
King, David J.