Coupling mammalian cell surface display with somatic hypermutation for the discovery and maturation of human antibodies

被引：81

作者：

Bowers, Peter M. ^{[1
]}

Horlick, Robert A. ^{[1
]}

Neben, Tamlyn Y. ^{[1
]}

Toobian, Rachelle M. ^{[1
]}

Tomlinson, Geoffrey L. ^{[1
]}

Dalton, Jennifer L. ^{[1
]}

Jones, Heather A. ^{[1
]}

Chen, Andy ^{[1
]}

Altobell, Laurence, III ^{[1
]}

Zhang, Xue ^{[1
]}

Macomber, John L. ^{[1
]}

Krapf, Irina P. ^{[1
]}

Wu, Betty F. ^{[1
]}

McConnell, Audrey ^{[1
]}

Chau, Betty ^{[1
]}

Holland, Trevin ^{[1
]}

Berkebile, Ashley D. ^{[1
]}

Neben, Steven S. ^{[1
]}

Boyle, William J. ^{[1
]}

King, David J. ^{[1
]}

机构：

[1] AnaptysBio Inc, San Diego, CA 92121 USA

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2011年 / 108卷 / 51期

关键词：

affinity maturation; mammalian display; NERVE GROWTH-FACTOR; HUMAN MONOCLONAL-ANTIBODIES; V-LAMBDA REPERTOIRE; MEMORY B-CELLS; AFFINITY MATURATION; RAPID GENERATION; DIVERSITY; EVOLUTION; GENE; LINE;

D O I：

10.1073/pnas.1114010108

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

070301 [无机化学]; 070403 [天体物理学]; 070507 [自然资源与国土空间规划学]; 090105 [作物生产系统与生态工程];

摘要：

A novel approach has been developed for the isolation and maturation of human antibodies that replicates key features of the adaptive immune system by coupling in vitro somatic hypermutation (SHM) with mammalian cell display. SHM is dependent on the action of the B cell specific enzyme, activation-induced cytidine deaminase (AID), and can be replicated in non-B cells through expression of recombinant AID. A library of human antibodies, based on germline V-gene segments with recombined human (D) J regions was used to isolate low-affinity antibodies to human beta nerve growth factor (h beta NGF). These antibodies, initially naive to SHM, were subjected to AID-directed SHM in vitro and selected using the same mammalian cell display system, as illustrated by the maturation of one of the antibodies to low pM K(D). This approach overcomes many of the previous limitations of mammalian cell display, enabling direct selection and maturation of antibodies as full-length, glycosylated IgGs.

引用

页码：20455 / 20460

页数：6

共 42 条

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