L&T Ocular Pathology Department, Vision Research Foundation, Sankara Nethralaya, No. 41, College Road, Nungambakkam, Chennai, Tamil Nadu, 600006, India.
School of Chemical and Biotechnology, SASTRA Deemed University, Thanjavur, India.
Sci Rep. 2024 Jun 12;14(1):13552. doi: 10.1038/s41598-024-64559-7.
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.
抗体在基于蛋白质组学的技术的众多应用中被广泛使用，需要高灵敏度、特异性、宽动态检测范围以及精确、可重复的定量。由于抗体存在一些固有局限性，因此寻找替代抗体的方法是一个非常重要的研究领域。最近，适体越来越受到关注，因为它们不仅具有抗体的所有优势，而且具有独特的优势，如热稳定性、低成本和无限的应用。适体在免疫学研究中越来越重要，并且有可能在免疫测定中替代抗体。B7H3 是一种免疫调节蛋白，属于 B7 家族，由于其在几种肿瘤组织中过度表达，而在正常组织中表达有限，因此是一个有吸引力和有前途的靶标。本研究采用杂交-SELEX 与下一代测序相结合的方法，筛选出特异性结合 B7H3 蛋白的 ssDNA 适体。这些适体在包括流式细胞术、斑点印迹和免疫组织化学在内的各种检测中表现出多功能性。在夹心斑点印迹和 Western blot 分析中的有效性能表明它们具有用于诊断应用的潜力，并证明了它们在各种蛋白质检测技术中的适应性和成本效益。
