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使用酶联免疫吸附测定法对眼镜王蛇毒液神经毒素、磷脂酶A2、出血毒素和L-氨基酸氧化酶的抗原交叉反应性进行研究。

An investigation into the antigenic cross-reactivity of Ophiophagus hannah (king cobra) venom neurotoxin, phospholipase A2, hemorrhagin and L-amino acid oxidase using enzyme-linked immunosorbent assay.

作者信息

Tan N H, Lim K K, Jaafar M I

机构信息

Department of Biochemistry, University of Malaya, Kuala Lumpur.

出版信息

Toxicon. 1993 Jul;31(7):865-72. doi: 10.1016/0041-0101(93)90221-4.

Abstract

The antigenic cross-reactivity of four Ophiophagus hannah (king cobra) venom components, the neurotoxin (OH-NTX), phospholipase A2 (OH-PLA2), hemorrhagin (OH-HMG) and L-amino acid oxidase (OH-LAAO) were examined by indirect and double sandwich ELISAs. The indirect ELISAs for OH-NTX, OH-PLA2 and OH-HMG were very specific when assayed against the various heterologous snake venoms and O. hannah venom components, at 25 ng/ml antigen level. At higher antigen concentrations (100-400 ng/ml), there were moderate to strong indirect ELISA cross-reactions between anti-O. hannah neurotoxin and venoms from various species of cobra as well as two short neurotoxins. However, anti-O. hannah hemorrhagin did not cross-react with any of the venoms tested, even at these high antigen concentrations, indicating that O. hannah hemorrhagin is antigenically very different from other venom hemorrhagins. Examination of the indirect ELISA cross-reactions between anti-O. hannah PLA2 and several elapid PLA2 enzymes suggests that the elapid PLA2 antigenic class has more than two subgroups. The antibodies to O. hannah L-amino acid oxidase, however, yielded indirect ELISA cross-reactions with many venoms as well as with OH-NTX, OH-PLA2 and OH-HMG, indicating that OH-LAAO shares common epitopes even with unrelated proteins. The double sandwich ELISAs for the four anti-O. hannah venom components, on the other hand, generally exhibited a higher degree of selectivity than the indirect ELISA procedure.

摘要

通过间接酶联免疫吸附测定法(ELISA)和双夹心ELISA检测了眼镜王蛇(Ophiophagus hannah)四种毒液成分,即神经毒素(OH-NTX)、磷脂酶A2(OH-PLA2)、出血毒素(OH-HMG)和L-氨基酸氧化酶(OH-LAAO)的抗原交叉反应性。当以25 ng/ml的抗原水平针对各种异源蛇毒和眼镜王蛇毒液成分进行检测时,针对OH-NTX、OH-PLA2和OH-HMG的间接ELISA具有高度特异性。在较高抗原浓度(100 - 400 ng/ml)下,抗眼镜王蛇神经毒素与各种眼镜蛇毒液以及两种短神经毒素之间存在中度至强烈的间接ELISA交叉反应。然而,即使在这些高抗原浓度下,抗眼镜王蛇出血毒素也未与任何测试毒液发生交叉反应,这表明眼镜王蛇出血毒素在抗原性上与其他毒液出血毒素有很大差异。对抗眼镜王蛇PLA2与几种眼镜蛇科PLA2酶之间的间接ELISA交叉反应进行检测表明,眼镜蛇科PLA2抗原类别有两个以上的亚组。然而,抗眼镜王蛇L-氨基酸氧化酶抗体与许多毒液以及OH-NTX、OH-PLA2和OH-HMG产生了间接ELISA交叉反应,这表明OH-LAAO即使与不相关的蛋白质也有共同的表位。另一方面,针对四种抗眼镜王蛇毒液成分的双夹心ELISA通常比间接ELISA方法表现出更高的选择性。

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