基于硫醇自组装的肌红蛋白单克隆抗体金膜固相载体的构建

Construction of gold film solid-phase carrier of myoglobin monoclonal antibody based on thiol self-assembly

摘要:

为了在金膜固相载体上固定肌红蛋白单克隆抗体 (MbAb)，通过在金膜上生长一层巯基酸和巯基醇的混合自组装分子膜 (SAMs)，用原子力显微镜 (AFM) 和X射线光电子能谱仪 (XPS) 分析样品的性质，再以1-(3-二甲基氨基丙基) 3-乙基碳化二亚胺盐酸 (EDC·HCl) 作为催化剂，自组装膜和抗体的氨基发生偶联反应，将抗体固定在金膜表面，并进行肌红蛋白抗原 (MbAg) 的测定。结果显示，通过条件优化实验，发现50 mmol/L的巯基16酸和巯基11醇混合乙醇溶液，60 ℃处理金膜3 h后，再偶联

Abstract:

In order to immobilize myoglobin (Mb) monoclonal antibody on gold film solid-phase carrier, we grew a mixed self-assembled monolayers (SAMs) of acid thiol and mercapto ethanol on gold film. Then we analyzed the property of the sample by atomic force microscopy and X-ray photoelectron spectroscopy. Then, we used 1-(3-dimethyl aminopropyl)-3-ethyl carbodiimide hydrochloride as catalyst to couple SAMs with amino of antibody so that we immobilized antibody on surface of gold film, followed by detecting myoglobin antigen. Results showed that, by optimizing experimental conditions, when we treated gold film by a mixture of mercapto hexadecanoic acid and mercapto undecanol ethanol solution of concentration of 50 mmol/L at temperature of 60 °C for 3 hours, and Mb monoclonal antibody of concentration of 40 mg/L for 3 hours, respectively, antibody had high immobilization efficiency and the MbAg was detected to 30 μg/L. The method provided a theoretical and practical basis for using magnetoresistence biosensors to diagnosis myocardial infarction.