Santoro F, Afchain D, Pierce R, Cesbron J Y, Ovlaque G, Capron A
Clin Exp Immunol. 1985 Nov;62(2):262-9.
The major surface protein (P30) of Toxoplasma gondii has been purified by immunoabsorption with anti-P30 monoclonal antibodies linked to a glutardialdehyde activated affinity absorbant. SDS-PAGE analysis of the eluted material followed by silver staining showed only a single band of 30,000 mol wt. Western blotting using antibodies from a rabbit immunized with purified P30 against the total Toxoplasma extract separated by SDS-PAGE again revealed an unique antigen of 30,000 daltons. The presence of repeated epitopes within P30 was confirmed by a two-site/one-antibody radiometric assay with the purified protein. Sandwich ELISA procedures with purified P30 clearly demonstrated that all 37 tested patients with acute toxoplasmosis presented significantly high levels of IgM anti-P30 antibodies. In addition, all 40 tested patients with chronic toxoplasma infection also showed high IgG anti-P30 antibody levels. These findings represent an essential step for the development of new reagents for the diagnosis of toxoplasmosis.
通过与连接到戊二醛活化亲和吸附剂上的抗P30单克隆抗体进行免疫吸附，已纯化了刚地弓形虫的主要表面蛋白（P30）。对洗脱物质进行SDS-PAGE分析，随后进行银染，结果显示只有一条分子量为30,000的条带。使用用纯化的P30免疫的兔子产生的抗体对经SDS-PAGE分离的刚地弓形虫总提取物进行蛋白质印迹分析，再次揭示了一种独特的30,000道尔顿抗原。用纯化蛋白进行的双位点/单抗体放射分析证实了P30内存在重复表位。用纯化的P30进行的夹心ELISA程序清楚地表明，所有37例接受检测的急性弓形虫病患者的抗P30 IgM抗体水平均显著升高。此外，所有40例接受检测的慢性弓形虫感染患者的抗P30 IgG抗体水平也很高。这些发现是开发用于诊断弓形虫病的新试剂的重要一步。
