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为制备纯化的灭活SARS病毒,取广东省CDC提供的灭活SARS-CoV病毒培养液进行超滤,收集相对分子质量介于100000至500000之间的组分,并浓缩。对纯化过程中各组分进行抗原活性鉴定和蛋白质含量测定,并加佐剂后免疫动物,测定抗体免疫应答。结果表明,超滤后活性抗原蛋白质回收率7.6%,纯化倍数为9,抗原活性达到1:480。免疫家兔获得的血清ELISA效价达到1:10000以上,中和抗体效价达到1:2000以上。表明该纯化工艺能获得较高纯度的灭活SARS-CoV病毒颗粒。
To prepare purified inactivated SARS virus, the inactivated SARS-CoV virus culture solution provided by CDC of Guangdong Province was ultrafiltrated, and the components with the relative molecular mass of between 100,000 and 500,000 were collected and concentrated. The components of the purification process for the identification of antigen activity and protein content determination, and adjuvants were immunized animals, antibody immune response was measured. The results showed that after ultrafiltration the active antigen protein recovery rate of 7.6%, the purification fold of 9, the antigen activity of 1: 480. The serum ELISA titer obtained by immune rabbits reached above 1: 10000, and the neutralizing antibody titer reached above 1: 2000. This shows that the purification process can obtain higher purity inactivated SARS-CoV virus particles.