Nabavinia Maryam Sadat, Charbgoo Fahimeh, Alibolandi Mona, Mosaffa Fatemeh, Gholoobi Aida, Ramezani Mohammad, Abnous Khalil
Department of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
Pharmaceutical Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
Appl Biochem Biotechnol. 2018 Feb;184(2):444-452. doi: 10.1007/s12010-017-2548-7. Epub 2017 Jul 19.
Aptamers are single-stranded RNA or DNA, which bind to their target with high affinity and specificity. Method of isolating aptamers against cell surface protein is called cell-SELEX. Common approach for monitoring cell-SELEX developed aptamers is flow cytometry. Since flow cytometry is costly and requires sophisticated equipments, we suggested implementing easy access, high throughput enzyme-link apta-sorbent assay test (ELASA) to confirm the specificity of aptamers selected through cell-SELEX process. In this regard, we compared ELASA and flow cytometry techniques in order to screen potent candidate aptamers against A2780 Rcis cell line, which were selected by cell-SELEX. The obtained results demonstrated that both ELASA and flow cytometry are identical in terms of sensivity and precision for aptamers selection. Then it could be concluded that ELASA method could be used as a versatile, inexpensive procedure for in vito evaluation of isolated aptamers from cell-SELEX based process.
适体是单链RNA或DNA，它们以高亲和力和特异性与其靶标结合。针对细胞表面蛋白分离适体的方法称为细胞指数富集的配体系统进化技术（cell-SELEX）。监测通过细胞指数富集的配体系统进化技术开发的适体的常用方法是流式细胞术。由于流式细胞术成本高昂且需要精密设备，我们建议采用易于操作、高通量的酶联适体吸附测定试验（ELASA）来确认通过细胞指数富集的配体系统进化技术过程筛选出的适体的特异性。在这方面，我们比较了酶联适体吸附测定试验和流式细胞术技术，以筛选针对通过细胞指数富集的配体系统进化技术选择的A2780 Rcis细胞系的有效候选适体。获得的结果表明，在适体选择的灵敏度和精度方面，酶联适体吸附测定试验和流式细胞术是相同的。因此可以得出结论，酶联适体吸附测定试验方法可以用作一种通用、廉价的程序，用于体外评估从基于细胞指数富集的配体系统进化技术的过程中分离出的适体。
