Microarrays with varying carbohydrate density reveal distinct subpopulations of serum antibodies.
Microarrays with varying carbohydrate density reveal distinct subpopulations of serum antibodies.
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DOI：
10.1021/pr9002245
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发表时间：
2009-07
影响因子：
4.4
通讯作者：
Gildersleeve JC
中科院分区：
文献类型：
--
作者：
Oyelaran O;Li Q;Farnsworth D;Gildersleeve JC
抗原阵列已成为分析蛋白质（如血清抗体）的复杂混合物的重要工具。这些阵列可用于更好地了解免疫反应，发现新的生物标志物，并指导疫苗的开发。然而，它们并不完美，改进的阵列设计将增强从这项技术中获得的信息。在这项研究中，我们描述和评估的战略，不同的抗原密度的阵列，然后使用该阵列来研究凝集素，单克隆抗体和血清抗体的结合。为了改变密度，合成了含有不同量碳水化合物的新糖蛋白，并用于制备结构和密度都不同的碳水化合物微阵列。我们证明，这种方法提供了一个范围内，是相关的生物识别事件的阵列表面上的密度的变化。该阵列用于评估三种凝集素（野豌豆凝集素B4、Happylopomatia凝集素和大豆凝集素）和三种结合肿瘤相关Tn抗原的单克隆抗体（HBTn-1、B1.1和Bric 111）的密度依赖性结合特性。此外，对30名健康供体的血清抗体进行了分析。结果表明，抗原密度的变化是检测结合特定抗原的抗体的全谱所必需的，并且可以用于揭示使用单一抗原密度不可检测的个体之间的抗体群体的差异。
Antigen arrays have become important tools for profiling complex mixtures of proteins such as serum antibodies. These arrays can be used to better understand immune responses, discover new biomarkers, and guide the development of vaccines. Nevertheless, they are not perfect and improved array designs would enhance the information derived from this technology. In this study, we describe and evaluate a strategy for varying antigen density on an array and then use the array to study binding of lectins, monoclonal antibodies, and serum antibodies. To vary density, neoglycoproteins containing differing amounts of carbohydrate were synthesized and used to make a carbohydrate microarray with variations in both structure and density. We demonstrate that this method provides variations in density on the array surface within a range that is relevant for biological recognition events. The array was used to evaluate density dependent binding properties of three lectins (Vicia villosa lectin B4, Helix pomatia agglutinin, and soybean agglutinin) and three monoclonal antibodies (HBTn-1, B1.1, and Bric111) that bind the tumor-associated Tn antigen. In addition, serum antibodies were profiled from 30 healthy donors. The results show that variations in antigen density are required to detect the full spectrum of antibodies that bind a particular antigen and can be used to reveal differences in antibody populations between individuals that are not detectable using a single antigen density.
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影响因子：
15
作者：
Gordus, Andrew;MacBeath, Gavin
通讯作者：
MacBeath, Gavin
影响因子：
4.8
作者：
ADLER, P;WOOD, SJ;SCHNAAR, RL
通讯作者：
SCHNAAR, RL
DOI：
10.1016/j.bbrc.2008.04.120
发表时间：
2008-07-11
影响因子：
3.1
作者：
Kato, Kentaro;Takeuchi, Hideyuki;Irimura, Tatsuro
通讯作者：
Irimura, Tatsuro
影响因子：
15
作者：
Gestwicki, JE;Cairo, CW;Kiessling, LL
通讯作者：
Kiessling, LL
DOI：
10.1073/pnas.94.20.10554
发表时间：
1997-09-30
影响因子：
11.1
作者：
Liang, R;Loebach, J;Kahne, D
通讯作者：
Kahne, D