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免疫分析方法主要依赖于固定在固相基底上的抗体对相应抗原的特异性识别,抗体在固相基底上固定后最大程度保持生物学活性是设计免疫分析方法的关键技术.本研究利用在疏水性硅基底表面制备蛋白A单分子膜层可以特异结合抗体的Fc端,进而实现对抗体的定向化装配,进一步构建格式化阵列用于肿瘤标志物检测.研究结果表明,制备的蛋白A单分子膜层的厚度为1.8±0.6nm,抗体可以经蛋白A单分子膜层实现定向化装配,由此设计的传感阵列检测肿瘤标志物甲胎蛋白(alpha-fetoprotein,AFP)可以实现1.0ng/mL的灵敏度,血清检测结果与电化学法(ECLIA)测定进行统计学检验没有显著性差异(P>0.05).
The immunoassay mainly depends on the specific recognition of the corresponding antigen by the antibody immobilized on the solid phase substrate, and the most important biological activity of the antibody immobilized on the solid phase substrate is the key technology for designing the immunoassay method.In this study, The monolayer of protein A on the surface of the silicon substrate can specifically bind to the Fc end of the antibody to realize the directed assembly of the antibody and to further construct a formatted array for tumor marker detection.The results show that the prepared protein A single molecule The thickness of the membrane layer is 1.8 ± 0.6nm, the antibody can be directed assembly through the protein A monolayer, and the designed sensor array can detect tumor marker alpha-fetoprotein (AFP) 1.0ng / mL sensitivity, there was no significant difference between the serum test results and the electrochemical test (ECLIA) for statistical test (P> 0.05).