Bertram M A, Griffiss J M, Broud D D
J Immunol. 1976 Mar;116(3):842-6.
Adaptations of the Farr technique have resulted in a specific and reproducible radioactive antigen-binding assay for antibodies directed against the lipopolysaccharide (LPS) of Neisseria meningitidis. LPS was intrinsically labeled with 14C acetate during 16-hr growth in a modified Frantz media, extracted by hot phenol-H2O, and purified by dialysis, ultracentrifugation, and ethanol precipitation. LPS, which aggregates in aqueous solutions, was maintained in a monomeric form in 3% sodium deoxycholate (NaD) as determined by gel filtration on Sephadex G-75. Since NaD is insoluble in (NH4)2SO4, polyethylene glycol, 20%, was used to precipitate immunoglobulins of all three major classes.
对法尔技术的改进已产生了一种针对脑膜炎奈瑟菌脂多糖(LPS)抗体的特异性且可重复的放射性抗原结合测定法。在改良的弗兰茨培养基中16小时生长期间,LPS用14C乙酸盐进行内在标记,通过热酚 - 水提取,并通过透析、超速离心和乙醇沉淀进行纯化。通过在Sephadex G - 75上进行凝胶过滤测定,在水溶液中聚集的LPS在3%脱氧胆酸钠(NaD)中保持单体形式。由于NaD不溶于硫酸铵((NH4)2SO4),因此使用20%的聚乙二醇沉淀所有三种主要类别的免疫球蛋白。