不同因素对酶联免疫法测定破伤风免疫球蛋白活性的影响

Effect of the different factors on the assay of tetanus immunoglobulin activity

目的考察不同因素对酶联免疫法（ELISA）测定破伤风免疫球蛋白生物活性的影响。方法将破伤风免疫球蛋白加入不同pH的缓冲液、不同浓度的氯化钠、硫酸铵以及聚乙二醇（PEG，相对分子质量为1000，6000和10000）溶液，采用ELISA法测定其活性。结果pH5～9的缓冲溶液对活性测定的影响不大，而pH≤4和≥10的缓冲溶液都会使测定值显著降低，当pH〈2或pH〉12时，活性下降到正常值的10％以下。随氯化钠浓度的增加测定值先下降到正常值的37．3％，后回升到41．2％，再下降；随硫酸铵浓度的增加测定值先下降到正常值的67％，后回升到99．4％，再下降。PEG对活性测定有较大影响，随PEG浓度的增加活性测定值降低。结论缓冲液pH、盐以及PEG对破伤风免疫球蛋白活性测定有影响。

Purpose The influencing factors in the activity determination of tetanus immunoglobulin using enzyme-linked immunosorbent assay （ELISA） were studied. Methods By adding tetanus immunoglobulin to buffer solutions of different pH, sodium chloride solutions, ammonia sulfate solutions and polyethylene glycol solutions （PEG, molecular weight ： 1 000,6 000, 10 000） of different concentrations, the activity was determined with ELISA. Results The results showed that pH 5-9 buffer solutions have no obvious influence on it, but in too acidic or too alkaline media the activity measure values are abased. The effect of sodium chloride and ammonia sulfate solution is complex in these solutions with the rising of the concentration, the activity declined gradually, then ascended, finally declined again. The ELISA result was reduced as PEG was added in. Additionally, the smaller PEG molecular weight is, the more markedly the infection is. Conclusion The ELISA results are influenced by different pH buffer solutions, salt and PEG.