Antibody-bound cell microarray for immunophenotyping: surface modification and lymphocyte subpopulations.
Antibody-bound cell microarray for immunophenotyping: surface modification and lymphocyte subpopulations.
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用于免疫表型分析的抗体结合细胞微阵列：表面修饰和淋巴细胞亚群。
DOI：
10.1002/jbm.b.31139
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发表时间：
2008
期刊：
影响因子：
--
通讯作者：
Matsuda,Takehisa
中科院分区：
文献类型：
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作者：
Fujii,Yasuo;Anderson,JamesM;Matsuda,Takehisa
基于抗体的细胞微阵列可以作为一种高通量的诊断工具，它需要精确的表面设计，提供高生物学特异性、高可靠性和高有效性。用固定了血细胞分化簇(CD)抗原的抗体的细胞微阵列进行了免疫表型定量评价的基础研究。该微阵列是通过光微图案化烷基硫醇自组装单分子膜制备的，它由规则分布在甲基(1-十二硫醇)填充基质相中的羧基(3-巯基丙酸)填充结构域组成。X射线光电子能谱和水的润湿性测量证实了这一点。通过水溶性碳二亚胺和羟基琥珀酰亚胺的偶联反应，将直径为1.0 mm或0.2 mm的图案化羧化结构域与抗CD4或抗CD8抗体共价固定。通过激光共聚焦扫描显微镜结合特定的染料染色技术，验证了抗体在几乎完全转化时的精确固定。表达CD4和CD8抗原的外周血单核细胞分别以非常高的特异性附着在芯片的抗CD4或抗CD8抗体固定区域上，这一点通过流式细胞仪分析和抗体包裹的磁珠细胞分离技术得到了证实。用抗体固定芯片测定的CD4/CD8亚群比率与用流式细胞仪测定的结果非常接近。这些结果表明，细胞表面抗原特异性抗体共价固定的微阵列为高通量定量评估免疫表型在医学诊断中的应用提供了坚实的基础。#2008威利期刊公司生物材料资源B部分：APPL Biomater，2008
Antibody‐based cell microarrays may serve as a high‐throughput diagnostic tool that requires precise surface design providing high biological specificity, high reliability, and high validity. A fundamental study on the quantitative evaluation of immunophenotyping using a cell microarray on which antibodies specific to cluster of differentiation (CD) antigens of blood cells are fixed was performed. The microarray, which was prepared by photomicropatterning self‐assembled monolayers of alkanethiols, consisted of carboxyl group (3‐mercaptopropionic acid)‐packed domains regularly distributed in the methyl group (1‐dodecanethiol)‐packed matrix phase. This was verified by X‐ray photoelectron spectroscopy and water wettability measurements. The patterned carboxylated domains of 1.0 mm or 0.2 mm diameter were covalently fixed with the anti‐CD4 or anti‐CD8 antibody by coupling reaction using a water‐soluble carbodiimide and hydroxysuccinimide. Precision antibody fixation at almost complete conversion was verified by confocal laser scanning microscopy coupled with a specific dye staining technique. Peripheral blood mononuclear cells expressing CD4 or CD8 antigen, respectively, adhered on the anti‐CD4 or anti‐CD8 antibody‐fixed domains of the microarray at a very high specificity, which was verified with flow cytometric analysis and an antibody‐coated magnetic‐bead‐based cell isolation technique. The CD4/CD8 subset ratios determined using the antibody‐fixed microarrays were very close to those obtained by flow cytometric analysis. These results indicate that microarrays, on which antibodies specific to cell surface antigens are covalently fixed, provide a solid basis of the high‐throughput quantitative evaluation of immunophenotyping in medical diagnosis. © 2008 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2008