Data_Sheet_1_Novel ssDNA Ligand Against Ovarian Cancer Biomarker CA125 With Promising Diagnostic Potential.docx

The in-vitro diagnostic industry is always striving to explore specific and high-affinity recognition entities, sensitive probes, and newer technology or platforms to develop disease detection methods with lower production and time costs and with minimum interference or variability. Aptamers suffice as a reliable recognition element by addressing the issues mentioned earlier. Hence, this work focuses on screening high-affinity ssDNA ligands to capture an exemplary biomarker CA125 using membrane-SELEX technology coupled with aptainformatics (translational bioinformatics using aptamers). The ssDNA ligands or aptamers have been screened and characterized extensively through an array of assays to ensure a valuable diagnostic potential with KD (dissociation constant) of 166 nM. The robustness of the selected aptamer ligand 2.26 and its complex with target CA125 is investigated in the presence of serum and extreme salt concentrations. Its diagnostic potential is convincingly demonstrated by running a competitive nucleic acid lateral flow assay at various sample concentrations. The ssDNA ligand reported in this manuscript holds immense potential in the detection and specific targeting of CA125 biomarker.

体外诊断（in-vitro diagnostic）行业始终致力于探索特异性优异、亲和力强劲的识别分子、灵敏探针以及新兴技术与平台，以期开发出兼具更低生产成本与时间成本、且干扰性与变异性极低的疾病检测方法。适配体（Aptamers）可作为可靠的识别元件，解决上述提及的技术难题。因此，本研究依托膜SELEX（membrane-SELEX）技术结合适配体信息学（aptainformatics，即基于适配体的转化生物信息学），旨在筛选可捕获典型生物标志物CA125的高亲和力单链DNA（ssDNA）配体。研究团队通过一系列实验分析对筛选得到的ssDNA配体（即适配体）进行了全面表征，证实其具备极具应用价值的诊断潜力，其解离常数（KD, dissociation constant）为166 nM。本研究针对筛选得到的适配体配体2.26及其与靶标CA125形成的复合物，在血清环境与极端盐浓度条件下开展了稳定性测试以评估其稳健性。通过在不同样本浓度下开展竞争性核酸侧向流检测（competitive nucleic acid lateral flow assay），该适配体的诊断潜力得到了充分验证，结果令人信服。本文报道的ssDNA配体在CA125生物标志物的检测与特异性靶向领域具备极高应用潜力。