Directed evolution of high-affinity antibody mimics using mRNA display

被引：194

作者：

Xu, LH ^{[1
]}

Aha, P ^{[1
]}

Gu, K ^{[1
]}

Kuimelis, RG ^{[1
]}

Kurz, M ^{[1
]}

Lam, T ^{[1
]}

Lim, AC ^{[1
]}

Liu, HX ^{[1
]}

Lohse, PA ^{[1
]}

Sun, L ^{[1
]}

Weng, S ^{[1
]}

Wagner, RW ^{[1
]}

Lipovsek, D ^{[1
]}

机构：

[1] Phylos Inc, Lexington, MA 02421 USA

来源：

CHEMISTRY & BIOLOGY | 2002年 / 9卷 / 08期

关键词：

D O I：

10.1016/S1074-5521(02)00187-4

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We constructed a library of >10(12) unique, covalently coupled mRNA-protein molecules by randomizing three exposed loops of an immunoglobulin-like protein, the tenth fibronectin type III domain ((10)Fn3). The antibody mimics that bound TNF-alpha were isolated from the library using mRNA display. Ten rounds of selection produced (10)Fn3 variants that bound TNF-alpha with dissociation constants (K-d) between 1 and 24 nM. After affinity maturation, the lowest K-d measured was 20 pM. Selected antibody mimics were shown to capture TNF-alpha when immobilized in a protein microarray. (10)Fn3-based scaffold libraries and mRNA-display allow the isolation of high-affinity, specific antigen binding proteins; potential applications of such binding proteins include diagnostic protein microarrays and protein therapeutics.

引用

页码：933 / 942

页数：10