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抗草原蝰蛇毒神经生长因子的单克隆抗体与其他蛇毒神经生长因子发生交叉反应。

Monoclonal antibodies against Vipera lebetina venom nerve growth factor cross-react with other snake venom nerve growth factors.

作者信息

Arumäe U, Siigur J, Neuman T, Saarma M

机构信息

Department of Molecular Genetics, Academy of Sciences of the Estonian SSR.

出版信息

Mol Immunol. 1987 Dec;24(12):1295-302. doi: 10.1016/0161-5890(87)90124-6.

Abstract

Nerve growth factor (NGF) was isolated from the venom of Vipera lebetina and was purified to homogeneity as judged by SDS gel electrophoresis. The biologically active NGF was used to immunize BALB/c mouse, and the spleen cells from immunized mouse were fused with mouse PAI myeloma cells. Forty-seven hybrid cell lines, secreting monoclonal antibodies to V. lebetina NGF, were isolated and nine of them purified from ascitic fluids. The isolated antibodies define two partially overlapping epitopes of the V. lebetina NGF which are not involved in the biological activity of the molecule. Both epitopes are also present on the beta-NGF from the mouse salivary gland and on the NGFs from the following snake venoms: V. lebetina, V, ursini, V, berus berus, Echis carinatus, Bungarus caeruleus, Agkistrodon halys, Naja naja oxiana, Naja naja atra and Naja naja, but not on the bovine seminal plasma NGF. The mol. wts of the NGFs in these snake venoms were determined by Western immunoblot with monoclonal antibodies. The mol. wts of the NGFs from V. ursini (37,000), E. carinatus (36,000, 44,000) and A. halys (29,000) were determined for the first time.

摘要

神经生长因子(NGF)是从黎凡特蝰蛇的毒液中分离出来的,并通过SDS凝胶电泳判断其已纯化至同质。将具有生物活性的NGF用于免疫BALB/c小鼠,然后将免疫小鼠的脾细胞与小鼠PAI骨髓瘤细胞融合。分离出47个分泌针对黎凡特蝰蛇NGF的单克隆抗体的杂交细胞系,并从腹水液中纯化出其中9个。所分离的抗体确定了黎凡特蝰蛇NGF的两个部分重叠的表位,这些表位不参与该分子的生物活性。这两个表位也存在于小鼠唾液腺的β-NGF以及以下蛇毒的NGF上:黎凡特蝰蛇、乌氏蝰蛇、贝氏蝰蛇、锯鳞蝰蛇、蓝环蛇、白眉蝮蛇、中亚眼镜蛇、舟山眼镜蛇和眼镜蛇,但不存在于牛精浆NGF上。用单克隆抗体通过Western免疫印迹法测定了这些蛇毒中NGF的分子量。首次测定了乌氏蝰蛇(37,000)、锯鳞蝰蛇(36,000、44,000)和白眉蝮蛇(29,000)的NGF的分子量。

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