Sugar-binding proteins from fish: selection of high affinity "lambodies" that recognize biomedically relevant glycans.
Sugar-binding proteins from fish: selection of high affinity "lambodies" that recognize biomedically relevant glycans.
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DOI：
10.1021/cb300399s
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发表时间：
2013-01-18
影响因子：
4
通讯作者：
Pancer, Zeev
中科院分区：
文献类型：
--
作者：
Hong, Xia;Ma, Mark Z.;Gildersleeve, Jeffrey C.;Chowdhury, Sudipa;Barchi, Joseph J., Jr.;Mariuzza, Roy A.;Murphy, Michael B.;Mao, Li;Pancer, Zeev
糖结合蛋白在基础研究和临床应用中具有重要意义，但对糖具有高亲和力和选择性的糖结合蛋白很难获得。在这里，我们描述了一种简便和具有成本效益的策略，以产生单克隆七鳃鳗抗体，称为lambodies，目标聚糖决定簇。我们筛选了酵母表面展示（YSD）七鳃鳗可变淋巴细胞受体（VLR）的克隆库，可以选择性地结合各种生物医学上重要的糖表位。这些糖缀合物包括肿瘤相关碳水化合物抗原（Tn和TFα）、刘易斯抗原（莱亚和LeX）、N-羟乙酰神经氨酸、广泛中和HIV抗体的靶标（poly-Man 9和HIV gp 120）以及糖蛋白无唾液酸-绵羊颌下粘蛋白（aOSM）和无唾液酸-人血型糖蛋白A（aGPA）。我们分离了以聚糖依赖性方式结合这些靶点的克隆，并且具有非常强的结合常数，例如，通过表面等离子体共振（SPR）测定，Man 9为6.2 nM，gp 120为44.7 nM。通过聚糖阵列分析显示，一种特定的层状抗体VLRB. aGPA. 23对血型H 3型三糖（BG-H3，Fucα1-2Galβ1-3GalNAcα）、aGPA和TFα（Galβ1-3GalNAcα）具有选择性，亲和力常数分别为0.2、1和8 nM。在人类组织微阵列中，这种lambody选择性地检测了14种不同类型癌症中的癌症相关碳水化合物抗原。它染色了27%的非小细胞肺癌（NSCLC）样本，其模式与患者生存率低相关。具有对聚糖的精致亲和力和选择性的Lambody可以在糖生物学和生物医学研究中找到无数用途。
Glycan-binding proteins are important for a wide variety of basic research and clinical applications, but proteins with high affnity and selectivity for carbohydrates are diffcult to obtain. Here we describe a facile and cost-effective strategy to generate monoclonal lamprey antibodies, called lambodies, that target glycan determinants. We screened a library of yeast surface-displayed (YSD) lamprey variable lymphocyte receptors (VLR) for clones that can selectively bind various biomedically important glycotopes. These glycoconjugates included tumor-associated carbohydrate antigens (Tn and TFα), Lewis antigens (LeA and LeX), N-glycolylneuraminic acid, targets of broadly neutralizing HIV antibodies (poly-Man9 and the HIV gp120), and the glycoproteins asialo-ovine submaxillary mucin (aOSM) and asialo-human glycophorin A (aGPA). We isolated clones that bind each of these targets in a glycan-dependent manner and with very strong binding constants, for example, 6.2 nM for Man9 and 44.7 nM for gp120, determined by surface plasmon resonance (SPR). One particular lambody, VLRB.aGPA.23, was shown by glycan array analysis to be selective for the blood group H type 3 trisaccharide (BG-H3, Fucα1-2Galβ1-3GalNAcα), aGPA, and TFα (Galβ1-3GalNAcα), with affnity constants of 0.2, 1, and 8 nM, respectively. In human tissue microarrays this lambody selectively detected cancer-associated carbohydrate antigens in 14 different types of cancers. It stained 27% of non-small cell lung cancer (NSCLC) samples in a pattern that correlated with poor patient survival. Lambodies with exquisite affnity and selectivity for glycans may find myriad uses in glycobiology and biomedical research.
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影响因子：
64.8
作者：
Pancer, Z;Amemiya, CT;Cooper, MD
通讯作者：
Cooper, MD
影响因子：
--
作者：
Rittenhouse-Diakun, K;Xia, Z;Roy, R
通讯作者：
Roy, R
影响因子：
2.7
作者：
Sakai, Keiko;Yuasa, Noriyuki;Fujita-Yamaguchi, Yoko
通讯作者：
Fujita-Yamaguchi, Yoko
DOI：
10.1016/j.str.2010.05.016
发表时间：
2010-09-08
期刊：
Structure (London, England : 1993)
影响因子：
--
作者：
Moulaei T;Shenoy SR;Giomarelli B;Thomas C;McMahon JB;Dauter Z;O'Keefe BR;Wlodawer A
通讯作者：
Wlodawer A
影响因子：
3
作者：
Fujitani, N;Liu, Y;Kimura, H
通讯作者：
Kimura, H