PDF(Pubmed)
Abstract：
Antibodies have been extensively used in numerous applications within proteomics-based technologies, requiring high sensitivity, specificity, a broad dynamic range for detection, and precise, reproducible quantification. Seeking alternatives to antibodies due to several inherent limitations of antibodies is an area of active research of tremendous importance. Recently, aptamers have been receiving increasing attention, because they not only have all of the advantages of antibodies, but also have unique advantages, such as thermal stability, low cost, and unlimited applications. Aptamers are gaining importance in immunological studies and can potentially replace antibodies in immunoassays. B7H3, an immunoregulatory protein belonging to the B7 family, is an attractive and promising target due to its overexpression in several tumor tissues while exhibiting limited expression in normal tissues. This study employed hybrid-SELEX with next-generation sequencing to select ssDNA aptamers specifically binding to the B7H3 protein. These aptamers demonstrated versatility across various assays, including flow cytometry, dot-blot, and immunohistochemistry. Effective performance in sandwich dot-blot assays and western blot analysis suggests their potential for diagnostic applications and demonstrates their adaptability and cost-effectiveness in diverse protein detection techniques.
摘要：
抗体已广泛用于基于蛋白质组学的技术中的许多应用。需要高灵敏度，特异性，检测的宽动态范围，精确，可重复定量。由于抗体的几个固有限制而寻求抗体的替代品是非常重要的活跃研究领域。最近,适体受到越来越多的关注,因为它们不仅具有抗体的所有优点，但也有独特的优势，如热稳定性，低成本,和无限的应用程序。适体在免疫学研究中越来越重要，并可能替代免疫测定中的抗体。B7H3，一种属于B7家族的免疫调节蛋白，由于其在几种肿瘤组织中的过表达而在正常组织中表现出有限的表达,因此是有吸引力和有希望的靶标。这项研究采用杂交SELEX和下一代测序来选择与B7H3蛋白特异性结合的ssDNA适体。这些适体在各种分析中表现出多功能性，包括流式细胞术,斑点印迹,和免疫组织化学。在夹心斑点印迹分析和蛋白质印迹分析中的有效性能表明了它们在诊断应用中的潜力，并证明了它们在多种蛋白质检测技术中的适应性和成本效益。
