抗O型口蹄疫病毒单克隆抗体相对亲和力的测定

Measurement of relative affinity for monoclonal antibody against type O foot-and-mouth disease

目的:对3株O型口蹄疫病毒单克隆抗体(4A9、4C3、9F12)的相对亲和力进行测定。方法:用纯化的O型口蹄疫病毒作为包被抗原建立一种检测方法,将3株单克隆抗体(4A9、4C3、9F12)制备腹水并纯化,并用建立的检测方法测定3株单克隆抗体(4A9、4C3、9F12)的相对亲和力。结果:检测方法中抗原、酶标抗体的最佳稀释度分别为1∶800和1∶10000,单抗的相对亲和力分别为:4A9约在5.242μg/ml(1∶600),4C3约在0.763μg/ml(1∶5000),9F12约在0.127μg/ml(1∶40000),即9F12>4C3>4A9。结论:用建立的ELISA方法测定了3株O型口蹄疫病毒单克隆抗体(4A9、4C3、9F12)的相对亲和力,将为进一步研究口蹄疫病毒而选取合适的单抗提供理论依据。

Objective：Tu determine the relative affinity of three monoelonal antibodies （4A9,4C3,9F12） against type O foot-and- mouth diseaae. Methods： A methed was developed in which serial dilution of purified antigen of type O FMDV was eoated. The monoelonal antibodies （4A9,4C3,9F12） were purified from the asceites of mice inoculated with the hybfidonas. Relative affinity of the purified monoelonal antibodties was determined with the indirect ELISA method. Results： The optimal dilution of the antigen and enzyme-labeled antibodies were 1：800 and 1 ： 10 000, respectively. The relalive affnity of the monoelonal antibodies （4A9,4C3,9F12） were 5. 242 μg/ml （ 1 ： 600 ）, 0. 763 μg/ml （ 1 ： 5 000） and 0. 127 μg/ml（ 1：40 000）, respectively, with the relative affnity of 9F12 〉 4C3 〉 4A9. Conclusion： Relative affinity of the monoclonal antibodlies （4A9,4C3,9F12） can be determined by ELISA, and it provides a theoretical foundation for the monoclonal antibody whieh is suitable for used in further studies of FMDV.