“桥抗体”对酶免疫分析的影响

THE INFLUENCE OF “BRIDGE-ANTIBODY” ON ELISA

在对 T-2毒素检测的 ELISA 研究中,T2-HS(T2-琥珀酸半酯)与 BSA 或 PLL联接,前者(T2-C_4-BSA)为免疫原,后者(T2-C_4-PLL)为检测抗原。在进行 ELISA 时,将 T2-C_4-PLL 包被于微板,并与兔抗 T-2抗体和标准 T-2样品一起保温。T-2抗体结合量用酶(HRP)标羊抗兔 IgG 与酶底物反应进行测定。在此情况下,我们不仅观察到抗 T-2抗体及抗 BSA 抗体,而且还有抗“-C_4-”抗体(即“桥抗体”)存在,同时发现“桥抗体”对T-2检测的灵敏度有明显干扰,甚至使 ELISA 不能进行。如以 T2-C_2-TT 代替 T2-C_4-PLL为检测抗原,由于“-C_4-”桥的排除,“桥抗体”的干扰也明显减弱。

In the study of ELISA for detection of T-2 toxin,T2-HS (T2-hemisuccinate) was conjugated to BSA or PLL,the former (T2-G_4-BSA) was defined as the immunogen and the latter (T2-C_4-PLL) as the analytic antigen.In the ELISA,T2-C_4-PLL was coated to a microtiter plate and incubated with rabbit anti-T-2 antibody and standard T-2 sample. The amount of anti-T-2 antibody bound to the plate was determined by reaction with goat anti-rabbit IgG-HRP and by subsequent reaction with the substrate.In this instance,we observed not only the anti-T-2 antibody and the anti-BSA antibody,but also the anti- “-C_4-”antibody (i.e.“bridge antibody”).At the same time,a significant interference on the sensitivity to detection of T-2 was found and even more ELISA could not be carry out.If T2-C_2-TT was used as analytic antigeninstead of T2-C_4-PLL,the interference of“bridge antibody”was evidently weakened because of the absence of“-C_4-”bridge.