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目的探索三步法纯化及获得雪旺细胞源神经营养蛋白的方法，为获得纯化蛋白进行单克隆抗体研究奠定基础。方法收集预损伤大鼠坐骨神经２００条，用酶消化结合差速粘附法纯化并收集其中雪旺细胞，将细胞超声粉碎、离心，收集上清液超滤，再经ｓｅｐｈａｄｅｘＧ１００层析，得到两个主峰，分段收集，将其中活性部分过高效液相色谱柱。结果得到三个主峰，相对分子质量分别为６８×１０３、５４×１０３和３０×１０３，用ＭＴＴ法测定生物活性，５４×１０３蛋白生物活性最大，经ＳＤＳＰＡＧＥ电泳证实基本为单一蛋白带。结论应用三步法同时各步采用合适方案是纯化雪旺细胞源神经营养蛋白的较好方法。
Objective To explore a three-step purification of Schwann cells derived from neurotrophic protein and protein, and to obtain the purified protein for monoclonal antibody study. Methods 200 preconditioned sciatic nerve of rats were collected and purified by enzyme digestion combined with differential adhesion method. Schwann cells were collected and purified. The cells were sonicated and centrifuged. The supernatants were collected by ultrafiltration and then by sephadexG-100 chromatography. Two main peaks, collected in stages, will be active part of the over-high performance liquid chromatography column. Results Three main peaks were obtained. The relative molecular masses were 68 × 103, 54 × 103 and 30 × 103, respectively. The bioactivity was determined by MTT method. The maximum bioactivity of 54 × 103 protein was confirmed by SDSPAGE electrophoresis. Conclusion It is a good method to purify Schwann cell-derived neurotrophic protein by using the three-step method and the appropriate step in each step.