Klausen J, Magnusson M, Andersen A B, Koch C
Statens Seruminstitut, Copenhagen, Denmark.
Scand J Immunol. 1994 Sep;40(3):345-9. doi: 10.1111/j.1365-3083.1994.tb03471.x.
Nine monoclonal antibodies were raised against purified protein derivative (PPD) of tuberculin in mice previously treated with Bacilli Calmette Guérin (BCG). The antibodies also reacted with a culture filtrate from Mycobacterium tuberculosis strain H37Rv. In immunoblotting after SDS-PAGE the reaction with PPD was seen as a diffuse smear, whereas ammonium sulphate-precipitated proteins from H37Rv gave well-defined bands ranging from 10 to 65 kDa. Enzyme immunoassay showed that both PPD and H37Rv antigens were able to inhibit binding of the antibodies to PPD coated microtitre wells, suggesting that the antibodies reacted with continuous epitopes. A 12 kDa protein purified by immunoaffinity chromatography from H37Rv antigens was tested intradermally in M. tuberculosis MNC3 sensitized guinea pigs and gave a delayed type hypersensitivity reaction.
用卡介苗（BCG）预先处理过的小鼠，制备了9种抗结核菌素纯蛋白衍生物（PPD）的单克隆抗体。这些抗体也与结核分枝杆菌H37Rv菌株的培养滤液发生反应。在SDS-PAGE后的免疫印迹中，与PPD的反应表现为弥散的条带，而H37Rv菌株经硫酸铵沉淀的蛋白则产生了清晰的条带，范围在10至65 kDa之间。酶免疫测定表明，PPD和H37Rv抗原均能抑制抗体与包被有PPD的微量滴定孔的结合，提示这些抗体与连续表位发生反应。通过免疫亲和层析从H37Rv抗原中纯化出的一种12 kDa蛋白，在结核分枝杆菌MNC3致敏的豚鼠中进行皮内试验，引发了迟发型超敏反应。
