Costanzo Hayley, Gooch James, Frascione Nunzianda
King's College London, Department of Analytical, Environmental & Forensic Sciences London SE1 9NH UK
RSC Adv. 2025 Jul 1;15(28):22505-22523. doi: 10.1039/d5ra00645g. eCollection 2025 Jun 30.
Aptamers are short, single-stranded DNA or RNA oligonucleotides that can specifically bind to their target with high affinity and specificity. Aptamers have gained widespread attention in recent years as possible replacements for antibodies within many analytical fields, due to their high chemical and thermal stability and relative low cost of production. Red blood cells are of interest within not only the medical field, but also are of interest within forensic science. Few aptamers have been reported that can specifically detect human red blood cells, or surface proteins of, but they have great potential for use as biorecognition elements within immunoassays or biosensors. Three aptamers have been identified from recent literature that have been designed to bind to human red blood cells as a whole cell target or glycophorin A as a protein-based target. However, they are yet to be fully characterised for their binding affinity to red blood cells, and no sequence optimisation has been conducted. Within this work, a comprehensive characterisation of three reported aptamers has been conducted. modelling has been explored as a means to better understand the 3D structures and the target ligand of each aptamer. The 3D structures of these aptamers have been reported and utilised within the HDOCK server to predict the docking of the aptamers to red blood cell-specific surface proteins. Both enzyme-linked oligonucleotide assays and microscale thermophoresis have been used to characterise aptamer-target biding, with dissociation constants being predicted in the nanomolar to low micromolar range for each aptamer. Additionally, sequence optimisation has been conducted to enhance the binding of the sequences to human red blood cells through sequence truncation mechanisms. To the best of our knowledge, this work represents the first characterisation of these aptamers and will guide future use of these aptamers as analytical probes.
适体是短的单链DNA或RNA寡核苷酸，能够以高亲和力和特异性与它们的靶标特异性结合。近年来，由于其高化学和热稳定性以及相对较低的生产成本，适体作为许多分析领域中抗体的可能替代品而受到广泛关注。红细胞不仅在医学领域受到关注，在法医学领域也备受关注。很少有适体被报道能够特异性检测人类红细胞或其表面蛋白，但它们在免疫测定或生物传感器中作为生物识别元件具有巨大的应用潜力。从最近的文献中鉴定出三种适体，它们被设计用于结合作为全细胞靶标的人类红细胞或作为基于蛋白质的靶标的血型糖蛋白A。然而，它们与红细胞的结合亲和力尚未得到充分表征，也没有进行序列优化。在这项工作中，对三种报道的适体进行了全面表征。已经探索了建模作为更好地理解每种适体的三维结构和靶标配体的一种方法。这些适体的三维结构已经被报道，并在HDOCK服务器中用于预测适体与红细胞特异性表面蛋白的对接。酶联寡核苷酸测定和微量热泳动都已用于表征适体 - 靶标结合，预测每种适体的解离常数在纳摩尔到低微摩尔范围内。此外，已经通过序列截短机制进行了序列优化，以增强序列与人类红细胞的结合。据我们所知，这项工作代表了对这些适体的首次表征，并将指导这些适体作为分析探针的未来应用。
