大熊猫IgG双抗体夹心ELISA定量检测方法的建立

Development of double-antibody sandwich ELISA for quantitative detection of IgG in giant panda

为建立大熊猫IgG的定量双抗体夹心ELISA检测方法(DAS-ELISA),本研究纯化了大熊猫血清中IgG,并将其作为免疫原分别免疫家兔和豚鼠,制备兔抗和豚鼠抗大熊猫IgG高免血清,经优化DAS-ELISA反应条件,建立了以纯化的兔抗大熊猫IgG作为捕获抗体和豚鼠抗大熊猫IgG高免血清作为检测大熊猫IgG的定量DAS-ELISA检测方法。该方法能检测1.5625μg/mL～100μg/mL的大熊猫IgG,大熊猫IgG含量(Y)与DAS-ELISA检测OD值(X)之间呈良好的线性关系,Y=4.7186X+0.044,相关系数为0.9911,该方法特异性强,稳定性好,能用于大熊猫IgG的定量检测。

To establish a quantitative method for detecting giant panda IgG. Double-antibody sandwich ELISA （DAS-ELISA） was developed and optimized by using purified rabbit anti-giant panda IgG as capture antibody and guinea pig anti-giant panda IgG hyperimmune serum as detection antibody. The detection limits of the DAS-ELISA were between 1.5625 μg/mL to 100 μg/mL IgG per milliliter sera. A linear relationship between the giant panda IgG level ＂Y＂ and DAS-ELISA detection optical density （OD） ＂X＂ was established based on formula Y=4.7186X＋0.044, with the coefficient of determination （R^2） at 0.9911. The DAS-ELISA method was specific, stable and could be used in the quantitative detection of giant panda IgG.