小分子化合物多克隆抗体纯化方法优化及比较研究

Optimization and comparison of polyclonal antibody purification strategies

目的比较多克隆抗体的纯化方法,探讨多克隆抗体纯化的优化方案。方法以丙烯酰胺多克隆抗体为纯化对象,进行抗体浓度、识别特征、亲和常数等对比研究,比较盐析(辛酸-饱和硫酸铵法)、蛋白G亲和纯化、载体蛋白亲和纯化、全抗原亲和纯化4种抗体纯化方法对于小分子多克隆抗体的纯化效果。结果采用的4种方法对于多克隆抗体的纯化效果有一定差异,蛋白G亲和纯化与载体蛋白纯化联合使用的纯化效果较为理想,获得的抗体纯度高、单一识别小分子抗原,且具有最佳的亲和常数。结论通过本文研究,优化的蛋白G亲和纯化与载体蛋白纯化的组合方案提高了多克隆抗体质量,为后续免疫分析方法研究提供技术基础。

Objective To compare different antibody purification methods and establish an optimized strategy in order to obtain polyelonal antibodies with high purity and affinity. Methods Anti - acrylamide polyclonal antibody was used to evaluate four purification methods, including caprylic acid - saturated ammonium sulfate precipitation method, protein G purification method, carrier protein immuno - affinity purification method and complete antigen immuno - affinity purification method. And the resulted concentration, specificity and affinity of the obtained antibody were carefully studied. Results These four methods performed differently in polyclonal antibody purification. The combination of protein G purification and carrier protein immuno -affinity purification gave the most satisfactory results, providing the antibody with the highest purity, specificity and immunodfinity. Conclusion The optimized combination method for polyclonal antibody purification is established, enhancing the quality of obtained antibody and facilitating subsequent immunoassay research.