Ibsen P, Heron I
Vaccine Department, State Serum Institute, Copenhagen S. Denmark.
Biologicals. 1990 Apr;18(2):123-6. doi: 10.1016/1045-1056(90)90022-r.
The quantitation of pertussis toxin (PT) in two sandwich ELISAs was tested for specificity. The detection of the captured PT was obtained by using either polyspecific rabbit anti Bordetella pertussis serum (RaBp-ELISA) or a monoclonal anti-PT antibody (McaPT-ELISA). No major differences in the estimation of PT in highly purified preparations were noted using either ELISA variants. In contrast, the quantitation of PT in crude extracts of B. pertussis cultures by the RaBp-ELISA was found to be over-estimated and showed greater variability when compared to the McaPT-ELISA. Comparison of the distribution of PT in the eluate fractions following partial purification by hydroxylapatite chromatography revealed that the results of the McaPT-ELISA were more specific as judged by SDS-PAGE analysis.
对两种夹心酶联免疫吸附测定（ELISA）中百日咳毒素（PT）的定量进行了特异性测试。通过使用多特异性兔抗百日咳博德特氏菌血清（RaBp - ELISA）或单克隆抗PT抗体（McaPT - ELISA）来检测捕获的PT。使用任何一种ELISA变体，在高度纯化制剂中PT的估计值均未发现重大差异。相比之下，与McaPT - ELISA相比，发现通过RaBp - ELISA对百日咳博德特氏菌培养物粗提物中PT的定量被高估，且变异性更大。通过羟基磷灰石色谱法进行部分纯化后，对洗脱液组分中PT的分布进行比较，结果表明，通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳（SDS - PAGE）分析判断，McaPT - ELISA的结果更具特异性。
