Angenendt Philipp, Glökler Jörn, Murphy Derek, Lehrach Hans, Cahill Dolores J
Max Planck Institute for Molecular Genetics, Ihnestrasse 73, 14195 Berlin, Germany.
Anal Biochem. 2002 Oct 15;309(2):253-60. doi: 10.1016/s0003-2697(02)00257-9.
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 11 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.
随着蛋白质和抗体微阵列技术的出现，已经发表了几种不同的包被方法和实验方案，大致可分为两类：凝胶包被表面和普通的非凝胶包被的玻璃或塑料表面，有些带有连接的化学基团。我们筛选了这两种类型的11种不同的阵列表面，并比较了它们的检测限、芯片间和芯片内的差异以及储存特性。将五种不同的抗体固定在每种类型的微阵列载体上，每个点的总蛋白浓度范围为40飞摩尔至25阿托摩尔。从这些结果可以看出，有些抗体更适合用于抗体阵列。所有测量均重复进行四次，结果显示大多数普通玻璃和塑料载玻片具有高信号均匀性和可重复性。聚丙烯酰胺包被的载玻片检测限较低，使其更适合检测极低浓度的抗原。所有微阵列包被物均可储存8周；然而，储存2周后结果有所改善。总之，结果表明需要测试将用于抗体阵列的每种抗体，并根据实验要求选择微阵列包被物。
