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采用一种新的放射性抗原结合试验研究对脑膜炎奈瑟菌脂多糖抗原决定簇的反应。

Response to antigenic determinants of Neisseria meningitidis lipopolysaccharide investigated with a new radioactive antigen-binding assay.

作者信息

Bertram M A, Griffiss J M, Broud D D

出版信息

J Immunol. 1976 Mar;116(3):842-6.

PMID:56401
Abstract

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.

摘要

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。

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