Hilleman M R, McAleer W J, Buynak E B, McLean A A
Dev Biol Stand. 1983;54:3-12.
Preparation of hepatitis B vaccine in our laboratories consists of a series of steps that include initial concentration of surface antigen by ammonium sulfate precipitation, followed by isopycnic banding and rate zonal centrifugation in a K-II centrifuge. The partially purified antigen concentrate is digested with pepsin at pH2 and the antigen is unfolded in 8M urea solution followed by renaturation. After gel filtration, the antigen is treated with formalin in 1:4000 dilution, adsorbed onto alum, and preserved with thimerosal. The final product contains essentially pure hepatitis B surface antigen. The process relies both on physical elimination of infectious virus particles and treatment with highly viral-destructive reagents in the pepsin, urea and formalin steps. The process is known to be highly destructive of all known viruses tested and to include procedures that are known to be highly destructive of representatives of all known groups of animal viral agents. The three-step process in inactivation provides a fail-safe system for establishing safety of the product. Tests in more than 20'000 persons, who are under surveillance, have shown no untoward effect and have confirmed the safety of the product.
我们实验室制备乙肝疫苗包括一系列步骤，其中包括通过硫酸铵沉淀初步浓缩表面抗原，随后在K-II离心机中进行等密度梯度离心和速率区带离心。部分纯化的抗原浓缩物在pH2条件下用胃蛋白酶消化，抗原在8M尿素溶液中展开后再复性。经过凝胶过滤后，抗原用1:4000稀释的福尔马林处理，吸附到明矾上，并用硫柳汞保存。最终产品基本包含纯乙肝表面抗原。该过程既依赖于对感染性病毒颗粒的物理去除，也依赖于在胃蛋白酶、尿素和福尔马林步骤中使用高病毒破坏性试剂。已知该过程对所有测试的已知病毒具有高度破坏性，并且包括已知对所有已知动物病毒剂组的代表具有高度破坏性的程序。三步灭活过程为确定产品安全性提供了一个故障安全系统。对超过20000名受监测人员的测试表明没有不良影响，并证实了该产品的安全性。
