Dodd R Y, Kobita J A
J Immunol Methods. 1978;20:117-29. doi: 10.1016/0022-1759(78)90250-8.
Controlled pore glass (CPG) adsorbs hepatitis B surface antigen (HBsAg) from whole plasma with a high degree of specificity. The resultant complex is stable at acid pH and in the presence of high concentrations of sodium thiocyanate. The adsorbed HBsAg is qualitatively and quantitatively similar to the soluble material in its ability to bind antibodies to HBsAg (anti-HBs). The HBsAg in 1 ml of strongly reactive plasma is adsorbed by 100 mg of CPG, which can then specifically bind 32,000 passive hemagglutination units of anti-HBs. Bound antibody can be eluted in 77% yield by acid or by chaotropic ions and the CPG-HBsAg complex can be reused in further adsorption-elution cycles. Antibody to HBsAg can be purified 144-fold in a single step by using this technique. The preparation of monospecific subtyping reagents for HBsAg and of immunochemically purified anti-HBs is described.
可控孔径玻璃(CPG)能高度特异性地从全血浆中吸附乙型肝炎表面抗原(HBsAg)。所得复合物在酸性pH值及高浓度硫氰酸钠存在的情况下稳定。吸附的HBsAg在结合抗HBs抗体(anti-HBs)的能力方面,在质量和数量上与可溶性物质相似。1毫升强反应性血浆中的HBsAg可被100毫克CPG吸附,随后该CPG能特异性结合32,000个被动血凝单位的anti-HBs。结合的抗体可用酸或离液序列高的离子以77%的回收率洗脱,且CPG-HBsAg复合物可在进一步的吸附-洗脱循环中重复使用。使用该技术可在一步中把抗HBsAg抗体纯化144倍。本文描述了HBsAg单特异性分型试剂及免疫化学纯化的anti-HBs的制备方法。