ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, Future Industries Institute, University of South Australia, Mawson Lakes, South Australia 5095, Australia.
Experimental Therapeutics Laboratory, Hanson Institute and Samson Institute, School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, SA 5000, Australia.
Biosens Bioelectron. 2017 May 15;91:515-522. doi: 10.1016/j.bios.2017.01.004. Epub 2017 Jan 4.
A label-free porous silicon (pSi) based, optical biosensor, using both an antibody and aptamer bioreceptor motif has been developed for the detection of insulin. Two parallel biosensors were designed and optimised independently, based on each bioreceptor. Both bioreceptors were covalently attached to a thermally hydrosilylated pSi surface though amide coupling, with unreacted surface area rendered stable and low fouling by incorporation of PEG moieties. The insulin detection ability of each biosensor was determined using interferometric reflectance spectroscopy, using a range of different media both with and without serum. Sensing performance was compared in terms of response value, response time and limit of detection (LOD) for each platform. In order to demonstrate the capability of the best performing biosensor to detect insulin from real samples, an in vitro investigation with the aptamer-modified surface was performed. This biosensor was exposed to buffer conditioned by glucose-stimulated human islets, with the result showing a positive response and a high degree of selectivity towards insulin capture. The obtained results correlated well with the ELISA used in the clinic for assaying glucose-stimulated insulin release from donor islets. We anticipate that this type of sensor can be applied as a rapid point-of-use biosensor to assess the quality of donor islets in terms of their insulin production efficiency, prior to transplantation.
一种无标记的多孔硅（pSi）基光学生物传感器，同时使用抗体和适体生物受体基序，已被开发用于检测胰岛素。根据两种生物受体，设计并独立优化了两个平行的生物传感器。两种生物受体都通过酰胺偶联共价连接到热水解的 pSi 表面，通过引入 PEG 部分使未反应的表面区域稳定且不易受到污染。使用干涉反射光谱法，使用不同的介质（包括含血清和不含血清的介质），分别确定了每个生物传感器的胰岛素检测能力。从响应值、响应时间和检测限（LOD）等方面比较了两种平台的传感性能。为了证明性能最佳的生物传感器从实际样本中检测胰岛素的能力，对适体修饰表面进行了体外研究。将该生物传感器暴露于经葡萄糖刺激的人胰岛条件缓冲液中，结果显示对胰岛素的捕获具有阳性响应和高度选择性。获得的结果与临床上用于检测供体胰岛葡萄糖刺激胰岛素释放的 ELISA 相关性良好。我们预计，这种类型的传感器可以作为一种快速的现场使用生物传感器，用于评估供体胰岛的胰岛素产生效率，然后再进行移植。
