Cranfield Health, Cranfield University, Cranfield, UK.
Anal Chim Acta. 2012 Sep 5;741:1-8. doi: 10.1016/j.aca.2012.07.004. Epub 2012 Jul 10.
Within this paper we describe the use of scanning electrochemical microscopy (SECM) to fabricate a dotted array of biotinylated polyethyleneimine which was then used to immobilise first neutravidin and then a biotinylated antibody towards a relevant antigen of interest (PSA, NTx, ciprofloxacin). These antigens were selected both for their clinical relevance but also since they display a broad range of molecular weights, to determine whether the size of the antigen used effects the sensitivity of this approach. The SECM was then used to image the binding of both complementary and non-complementary antigens in a label-free assay. Imaging of the arrays before and following exposure to various concentrations of antigen in buffer showed clear evidence for specific binding of the complementary antigens to the antibody functionalised dots. Non-specific binding was also quantified by control experiments with other antigens. This demonstrated non-specific binding across the whole of the substrate, thereby confirming that specific binding does occur between the antibody and antigen of interest at the surface of the dots. The binding of ciprofloxacin was investigated both in simple buffer solution and in a more complex media, bovine milk.
在本文中，我们描述了扫描电化学显微镜（SECM）的使用，以制造出点状的生物素化聚乙烯亚胺阵列，然后将其用于固定首先是链亲和素，然后是针对相关感兴趣抗原（PSA、NTx、环丙沙星）的生物素化抗体。这些抗原既因其临床相关性而被选择，也因为它们显示出广泛的分子量范围，以确定所使用的抗原大小是否会影响这种方法的灵敏度。然后，SECM 用于在无标记测定中对互补和非互补抗原的结合进行成像。在缓冲液中暴露于不同浓度的抗原之前和之后对阵列进行成像，清楚地表明互补抗原与抗体功能化点之间存在特异性结合。通过用其他抗原进行对照实验也定量了非特异性结合。这证明了整个基底上的非特异性结合，从而证实了抗体和感兴趣的抗原确实在点的表面之间发生特异性结合。研究了环丙沙星在简单缓冲液和更复杂的介质（牛乳）中的结合情况。
