抗体对免疫传感界面的有效附着是决定免疫传感器检测性能的关键因素。因此,本研究旨在研究一种新型的低分子量抗体固定化材料,高稳定性,和优异的定向固定效果。在这项研究中,我们使用基于ZDOCK算法的分子对接技术来虚拟筛选抗体Fc段的DNA功能配体(DNAFL)。通过对DNAFL与IgG抗体界面的关键结合位点和接触倾向的综合分析,我们已经获得了对亲和力关系的宝贵见解,以及控制该界面上氨基酸和核苷酸相互作用的原理。此外,进行了分子亲和力实验和竞争性结合实验,以验证DNAFL与IgG抗体的结合能力及其实际结合位点。通过使用多碱基序列的亲和力实验,我们鉴定了显著影响抗体-DNAFL结合的碱基,并成功获得了对IgGFc片段亲和力增强的DNAFL.这些发现为高亲和力DNAFL的靶向设计提供了理论基础,同时也为免疫传感器的制备提供了新的技术方法,在生物检测中具有潜在的应用前景。
The effective attachment of antibodies to the immune sensing interface is a crucial factor that determines the detection performance of immunosensors. Therefore, this study aims to investigate a novel antibody immobilization material with low molecular weight, high stability, and excellent directional immobilization effect. In this study, we employed molecular docking technology based on the ZDOCK algorithm to virtually screen DNA functional ligands (DNAFL) for the Fc segment of antibodies. Through a comprehensive analysis of the key binding sites and contact propensities at the interface between DNAFL and IgG antibody, we have gained valuable insights into the
affinity relationship, as well as the principles governing amino acid and nucleotide interactions at this interface. Furthermore, molecular
affinity experiments and competitive binding experiments were conducted to validate both the binding ability of DNAFL to IgG antibody and its actual binding site. Through
affinity experiments using multi-base sequences, we identified bases that significantly influence antibody-DNAFL binding and successfully obtained DNAFL with an enhanced
affinity towards the IgG Fc segment. These findings provide a theoretical foundation for the targeted design of higher-
affinity DNAFLs while also presenting a new technical approach for immunosensor preparation with potential applications in biodetection.