Perez L, Estepa A, Coll J M
INIA-Sanidad Animal, CISA-Valdeolmos, Madrid, Spain.
J Virol Methods. 1998 Dec;76(1-2):1-8. doi: 10.1016/s0166-0934(98)00028-7.
A new method for the isolation of glycoprotein G from viral haemorrhagic septicaemia virus (VHSV), a fish rhabdovirus, was developed by using affinity chromatography with immobilized Concanavalin A (ConA). The glycoprotein G was isolated from detergent solubilized concentrated virions and from large-volume virion-free supernatants from VHSV infected cells (soluble form). The purity achieved was higher than 85%. The estimated recovery of the initial glycoprotein G present in the virions was between 20 and 50%. These glycoprotein G preparations showed the presence of about 30% of trimers by ultracentrifugation, reacted with antibodies to the phosphatidylserine binding domain (p2) in a pH-dependent manner by ELISA and bound phosphatidylserine in a pH-dependent manner by solid-phase binding assays. These data suggest that ConA purified glycoprotein G conserved most of its native properties and conformation.
一种从鱼类弹状病毒——病毒性出血性败血症病毒（VHSV）中分离糖蛋白G的新方法被开发出来，该方法使用了固定化伴刀豆球蛋白A（ConA）的亲和色谱法。糖蛋白G从经去污剂溶解的浓缩病毒粒子以及VHSV感染细胞的大量无病毒粒子上清液（可溶性形式）中分离得到。所达到的纯度高于85%。病毒粒子中初始糖蛋白G的估计回收率在20%至50%之间。通过超速离心分析，这些糖蛋白G制剂显示约30%为三聚体，通过酶联免疫吸附测定（ELISA），其与磷脂酰丝氨酸结合结构域（p2）的抗体以pH依赖的方式发生反应，并且通过固相结合测定以pH依赖的方式结合磷脂酰丝氨酸。这些数据表明，ConA纯化的糖蛋白G保留了其大部分天然特性和构象。
