Consiglio Nazionale delle Ricerche, Istituto di Chimica del Riconoscimento Molecolare (ICRM CNR), Milano, Italy.
J Immunol Methods. 2013 May 31;391(1-2):95-102. doi: 10.1016/j.jim.2013.02.013. Epub 2013 Feb 28.
The accuracy of antibody-based microarrays depends on eliminating or at least minimizing the effect of cross-reactive components. In this work, a method to create a versatile, compartmentalized storage system for antibodies and soluble microarray reagents is described. These containers are made of hydrogel plugs immobilized on a slide facing the corresponding sub-array on a printed surface. The hydrogels were polymerized using a 4% w/v N,N-dimethylacrylamide and 3% w/v bisacrylamide solution, thus generating large pore sizes to allow the facile transfer of intact, functional biomolecules. The hydrogel microenvironments can be desiccated and rehydrated with a desired solution, and they can store reagents in a dry form. These hydrogels were shown to provide limits of detection similar to those obtained by conventional incubation conditions in an assay for HIV p24 antigen. Moreover, they prevent cross-reactivity issues in the detection of Staphylococcus aureus enterotoxins A and B and promote the accurate quantification of interleukin 10 by a microarray-integrated calibration curve.
基于抗体的微阵列的准确性取决于消除或至少最小化交叉反应成分的影响。在这项工作中，描述了一种创建抗体和可溶性微阵列试剂的通用、分隔式存储系统的方法。这些容器由固定在打印表面相应子阵列上的滑动面上的水凝胶塞制成。水凝胶是使用 4%w/vN，N-二甲基丙烯酰胺和 3%w/v双丙烯酰胺溶液聚合的，从而产生大的孔径，以允许完整的、功能的生物分子的轻易转移。水凝胶微环境可以用所需的溶液进行干燥和再水合，并且可以以干燥形式储存试剂。这些水凝胶在 HIV p24 抗原检测的测定中显示出与传统孵育条件相似的检测限。此外，它们防止了金黄色葡萄球菌肠毒素 A 和 B 的检测中的交叉反应问题，并通过微阵列集成校准曲线促进了白细胞介素 10 的准确定量。
