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目的制备特异性敲除甘草酸的免疫亲和色谱柱。方法抗甘草酸单克隆抗体与CNBr活化的Sepharose 4BTM凝胶偶联制备甘草酸特异性敲除免疫亲和色谱柱,利用HPLC法测定敲除液与洗脱液中甘草酸质量浓度,考察甘草酸免疫亲和色谱柱的最大载样量、敲出率、精密度、准确度及稳定性等参数。结果甘草酸免疫亲和色谱柱的最大载样量为1.326 11 mg;日内及日间精密度总RSD分别为0.65%,相对误差(RE)为0.37%;免疫亲和色谱柱稳定性实验显示32 d内甘草酸敲除率的RSD为1.37%,稳定性良好。结论制备的甘草酸特异性敲除免疫亲和色谱柱能快速、高效、稳定地敲除甘草酸。
Objective To prepare immunoaffinity column with specific glycyrrhizin knockout. Methods Anti-glycyrrhizic acid monoclonal antibody was coupled with CNBr-activated Sepharose 4BTM gel to prepare glycyrrhizin-specific knockout immunoaffinity chromatography column. The mass concentration of glycyrrhizin in the knockout and eluate was determined by HPLC. Immunoaffinity column maximum load capacity, knockout rate, precision, accuracy and stability and other parameters. Results The maximum loading capacity of glycyrrhizin immunoaffinity column was 1.326 11 mg. The total RSD of intra-day and inter-day precision was 0.65% and the relative error (RE) was 0.37%. The stability of immunoaffinity column showed that The RSD of glycyrrhizin knockout rate in d was 1.37%, and the stability was good. Conclusion The glycyrrhizin-specific knockout immunoaffinity chromatography column can quickly and efficiently knock out glycyrrhizin.