Santucci A, Rustici M, Bracci L, Lozzi L, Soldani P, Neri P
CRISMA, Centro Didattico dell'Università, Siena, Italy.
J Immunol Methods. 1990 Feb 20;127(1):131-8. doi: 10.1016/0022-1759(90)90349-z.
It has been reported that the acetylcholine receptor may be used by the rabies virus to concentrate at sites in proximal to peripheral nerves. It has also been reported that the binding site for the receptor is located within the 190-203 region of the virus glycoprotein on the basis of its structural homology with the toxic center of snake neurotoxins, which are well known cholinergic ligands. We prepared monoclonal antibodies against the synthetic tetradecapeptide having the same sequence as the putative binding site of the rabies virus. One of three antibodies (clone 2PV 36-74) was able to recognize both the whole virus and its peplomeric glycoprotein and could bind acetylcholine. It was also able to inhibit the binding both of alpha-bungarotoxin and rabies virus glycoprotein to the acetylcholine receptor. We have covalently bound 2PV 36-74 to an HPLC affinity column and utilized it for specific purification of rabies virus glycoprotein. The immunoaffinity chromatographic method we describe is very sensitive and highly specific. Moreover this procedure does not denature the sample and is vary rapid and efficient.
据报道，狂犬病毒可能利用乙酰胆碱受体聚集在周围神经近端的部位。另据报道，基于其与蛇神经毒素毒性中心的结构同源性（蛇神经毒素是众所周知的胆碱能配体），该受体的结合位点位于病毒糖蛋白的190 - 203区域内。我们制备了针对与狂犬病毒假定结合位点具有相同序列的合成十四肽的单克隆抗体。三种抗体中的一种（克隆2PV 36 - 74）能够识别整个病毒及其包膜糖蛋白，并且能够结合乙酰胆碱。它还能够抑制α - 银环蛇毒素和狂犬病毒糖蛋白与乙酰胆碱受体的结合。我们已将2PV 36 - 74共价结合到HPLC亲和柱上，并将其用于狂犬病毒糖蛋白的特异性纯化。我们所描述的免疫亲和色谱方法非常灵敏且高度特异。此外，该程序不会使样品变性，而且快速高效。
