Pêtre J, Van Wijnendaele F, De Neys B, Conrath K, Van Opstal O, Hauser P, Rutgers T, Cabezon T, Capiau C, Harford N
SmithKline Biologicals, Rixensart, Belgium.
Postgrad Med J. 1987;63 Suppl 2:73-81.
The production in yeast cells of the recombinant DNA hepatitis B vaccine of SmithKline Biologicals involves an optimized fermentation process followed by cell disruption and extraction, together with other soluble yeast components of the surface antigen of the hepatitis B virus. The subsequent purification process includes precipitation steps, ion exchange and gel permeation chromatography, and caesium chloride ultracentrifugation. The yeast-derived antigen occurs as spherical particles containing the non-glycosylated HBsAg polypeptide, lipid, and Tween 20. The purity of the polypeptide is above 95% and confirmed by the absence of an immune response to yeast-derived contaminants in vaccinees. Yeast DNA levels were less than 10 pg/vaccine dose. Various biochemical analyses showed that the recombinant polypeptide was faithfully expressed and did not undergo unwanted processing or degradation during fermentation or purification. These results indicate that the recombinant HBsAg can be effectively produced in yeast and processed to a high degree of purity to yield HBsAg particles displaying most of the characteristic properties of plasma-derived HBsAg.
史克必成生物制品公司在酵母细胞中生产重组DNA乙型肝炎疫苗，涉及一个优化的发酵过程，随后进行细胞破碎和提取，以及乙肝病毒表面抗原的其他可溶性酵母成分。后续的纯化过程包括沉淀步骤、离子交换和凝胶渗透色谱法，以及氯化铯超速离心。酵母衍生的抗原以含有非糖基化HBsAg多肽、脂质和吐温20的球形颗粒形式存在。该多肽的纯度高于95%，并且通过疫苗接种者对酵母衍生污染物无免疫反应得到证实。酵母DNA水平低于10 pg/疫苗剂量。各种生化分析表明，重组多肽得到了忠实表达，在发酵或纯化过程中未发生不必要的加工或降解。这些结果表明，重组HBsAg可以在酵母中有效生产，并加工到高纯度，以产生具有血浆来源HBsAg大部分特性的HBsAg颗粒。
