Toward optimized antibody microarrays:: a comparison of current microarray support materials
Toward optimized antibody microarrays:: a comparison of current microarray support materials
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DOI：
10.1016/s0003-2697(02)00257-9
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发表时间：
2002-10-15
影响因子：
2.9
通讯作者：
Cahill, DJ
中科院分区：
文献类型：
--
作者：
Angenendt, P;Glökler, J;Cahill, DJ
随着蛋白质和抗体微阵列技术的出现，已经公布了几种不同的涂层和方案，其可以大致分为两种类型：凝胶涂覆的表面和普通的非凝胶涂覆的玻璃或塑料表面，一些表面附着有化学基团。我们已经筛选了II不同的阵列表面的两种类型，并比较它们的检测限，内部和芯片内的变化，和存储特性。将五种不同的抗体固定在每种类型的微阵列支持物上，总蛋白质浓度范围为40 fmol至25阿莫尔/点。从这些结果可以看出，一些抗体更适合用于抗体阵列。所有的测量都在四分之一次进行，结果显示大多数普通玻璃和塑料载玻片的高信号均匀性和再现性。用聚丙烯酰胺包被的载玻片获得较低的检测限，使其更适合检测极低浓度的抗原。所有微阵列涂层可储存8周;然而，储存2周后观察到改善的结果。总之，结果表明需要测试用于抗体阵列的每种抗体，并根据实验要求选择微阵列涂层。(C)2002 Elsevier Science（美国）。All rights reserved.
With the advent of protein and antibody microarray technology several different coatings and protocols have been published, which may be broadly divided into two types: gel-coated surfaces and plain non-gel-coated glass or plastic surfaces, some with chemical groups attached. We have screened I I different array surfaces of both types and compared them with respect to their detection limit, inter- and intrachip variation, and storage characteristics. Five different antibodies were immobilized onto each type of microarray support, with total protein concentrations ranging from 40 fmol to 25 amol per spot. From these results, it was seen that some antibodies were more suited for use on antibody arrays. All measurements were performed in quadruplicate, and the results revealed high signal uniformity and reproducibility of most plain glass and plastic slides. Lower detection limits were obtained with polyacrylamide-coated slides, making them more suitable for the detection of very low concentrations of antigen. All microarray coatings could be stored for a period of 8 weeks; however, improved results were seen after 2 weeks of storage. In conclusion, the results indicate the need to test each antibody to be used on an antibody array and to select the microarray coating based on experimental requirements. (C) 2002 Elsevier Science (USA). All rights reserved.