Coupling mammalian cell surface display with somatic hypermutation for the discovery and maturation of human antibodies
Coupling mammalian cell surface display with somatic hypermutation for the discovery and maturation of human antibodies
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DOI：
10.1073/pnas.1114010108
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发表时间：
2011-12-20
影响因子：
11.1
通讯作者：
King, David J.
中科院分区：
文献类型：
--
作者：
Bowers, Peter M.;Horlick, Robert A.;King, David J.
已经开发了一种用于分离和成熟人抗体的新方法，所述人抗体通过将体外体细胞超突变（SHM）与哺乳动物细胞展示偶联来复制适应性免疫系统的关键特征。SHM依赖于B细胞特异性酶、活化诱导的胞苷脱氨酶（AID）的作用，并且可以通过重组AID的表达在非B细胞中复制。基于具有重组人（D）J区的种系V基因片段的人抗体库用于分离针对人β神经生长因子（h β NGF）的低亲和力抗体。这些抗体，最初幼稚SHM，进行艾滋病定向SHM在体外和选择使用相同的哺乳动物细胞展示系统，如所示的成熟的抗体低pM K-D之一。这种方法克服了哺乳动物细胞展示的许多先前限制，使得能够直接选择抗体并使其成熟为全长糖基化IgG。
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.