【摘要】 在软化病毒抗血清制备中,开展了供免疫用抗原的简易纯化工作。主要步骤是将软化病蚕中肠在磷酸缓冲液及氯仿中抽提澄清,硫酸铵盐析,结合3000rpm离心30分钟,沉淀用少量pH7.2的磷酸缓冲液悬浮并经3000rpm离心60分钟,上清液即为供免疫用的纯抗原。用上法纯化所得病毒悬液,在紫外分光光度计上测得的吸收高峰为260,271nm;电子显微镜观察中除了球形病毒粒子外没有看到其他物质;用此法纯化为抗原所得到的抗血清在对流免疫电泳试验中特异性典型,且无交叉反应出现。更多还原

【Abstract】 In the production of antiserum against the flacherie virus, a simple process was developed for purifying the antigen for immunizing the rabbits. The process consisted essentially of extaracting the flacherie larva midguts in phosphate buffer, clarifying with chloroform, precipitating the virus with ammonium sulphate, followed then by centrifugation at 3000 rpm. for 30 min. The pellet obtained was suspended in small amount of phosphate buffer at pH7.2, and the suspension underwent 3000 rpm centrifugation for 60 min. The supernatant obtained was used as purified antigen for immunization. The virus suspension purified with this process showed,in ultraviolet spectrum, maximum absorption at 260,271am, showed spherical particles in electron micrograph with no noticeable extraneous material. Antiserum derived from the purified antigen showed specific reaction pattern in counter immuno-electrophoresis test, in which no cross-reactivity was observed.更多还原