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流感病毒血凝素和神经氨酸酶的分离提纯
Separation and Purification of Hemagglutinins and Neuraminidases of Influenza A Viruses
【摘要】 用甲醛化鸡红细胞吸附释放及葡聚糖G150柱层析,继之以高速离心沉淀制备纯化甲型流感病毒,纯度达3×10~5HAu/毫克蛋白。再经1~2%脱氧胆酸钠于37℃裂解后,用葡聚糖G150及DEAE纤维素(Cl-型)柱层析,经NaCl梯度洗脱,HA及NA分别洗脱于二个洗脱峰。圆盘电泳的结果表明,纯制的HA和NA在聚丙烯酰胺凝胶上于病毒的HA和NA各呈现单一的区带。
【Abstract】 Influenza A viruses were purified by means of adsorption and elution with formalinized chick RBC. After subjected to Sephadex G-150 chromato-graphy, they were further purified by high-speed centrifugation. The purified viruses were cleaved with 1-2% DOC for 30-60 minutes at 37℃and then subjected to another Sephadex G-150 column and eluted with 0.02M Tris-HC1 buffer, pH 8.0. Fractions containing hemagglutinin (HA) activity were pooled and concentrated with PEG until the total volumes were below 20 ml, then passed through a DEAE cellulose column (Cl-type) and eluted with NaCl gradients (0.1-0.8M) . Finally, the purified neuraminidase (NA) and HA were separated into two distinct peaks. Polyacrylamide gel electro-phoresis of the purified HA and NA showed that both preparations developed one single band on the gels.
- 【文献出处】 中国医学科学院学报 ,Acta Academiae Medicinae Sinicae , 编辑部邮箱 ,1982年01期
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