Bünger I, Khalaf H, Rimpler M
Staatlichen Veterinäruntersuchungsamt Hannover, Medizinischen Hochschule Hannover.
Dtsch Tierarztl Wochenschr. 1996 Dec;103(12):516-9.
8 different leukosis antigen preparations for immunodiffusion from 6 European countries were examined regarding their suitability for immunoblotting. Emphasis was made on demonstration of immunoreactions at 51 kDa beside reactions at 24 kDa. The whole protein content of the different preparations varied very much. gp 51 could be detected only by monoclonal antibodies at a molecular weight of 53 kDa, in one preparation at 26 kDa. With sera from field or with the European reference serum E4 none or only weak reactions could be observed at these molecular weights. All antigen preparations contained bovine IgG that is divided in L- and H-chains by SDS-electrophoresis under reducing conditions. These single chains reacted also with anti-bovine-IgG-biotin conjugate und disturbed the detection of the leukosis specific reactions. The bovine IgG could be removed by a Protein-G-column. One of all tested leukosis antigens, which are prepared for immunodiffusion, was suitable for immunoblotting. This antigen preparation contained only few bovine IgG and had strong reactions at 24 kDa. A special antigen preparation for detection of other reliable immunoreactions in the immunoblot has to be developed.
对来自6个欧洲国家的8种用于免疫扩散的白血病抗原制剂进行了检测,以评估它们是否适合免疫印迹法。重点是除了检测24 kDa处的免疫反应外,还要证明51 kDa处的免疫反应。不同制剂的总蛋白含量差异很大。gp 51仅在分子量为53 kDa时能被单克隆抗体检测到,在一种制剂中为26 kDa。用田间血清或欧洲参考血清E4在这些分子量处未观察到反应或仅观察到微弱反应。所有抗原制剂都含有牛IgG,在还原条件下通过SDS电泳可将其分为轻链和重链。这些单链也与抗牛IgG-生物素偶联物反应,并干扰了白血病特异性反应的检测。牛IgG可通过蛋白G柱去除。所有测试的用于免疫扩散的白血病抗原中,有一种适合免疫印迹法。这种抗原制剂仅含有少量牛IgG,在24 kDa处有强烈反应。必须开发一种用于在免疫印迹中检测其他可靠免疫反应的特殊抗原制剂。