Howard R J, Barnwell J W
Parasitology. 1984 Feb;88 ( Pt 1):27-36. doi: 10.1017/s0031182000054317.
Plasmodium knowlesi malaria-infected erythrocytes were radio-iodinated and several non-ionic, anionic and zwitterionic detergents were compared in their capacity to extract the labelled membrane proteins. The use of these detergents for antigen identification was tested by immunoprecipitation, after addition of Triton X-100 to some detergent extracts, using hyperimmune monkey antiserum and protein A-Sepharose. 125I-labelled antigens were specifically immunoprecipitated with all detergents tested, including the anionic detergents sodium dodecyl sulphate (SDS), deoxycholate and cholate; the zwitterions Zwittergent-312 and -314, CHAPS and Empigen BB, as well as several non-ionic detergents. The SDS-polyacrylamide gel electrophoresis patterns of 125I-labelled antigens varied after extraction with different detergents, there being no consistent pattern for detergents of a particular class. A total of 14 125I-labelled antigens were identified, 11 of them using Triton X-100. Some minor 125I-labelled antigens identified with Triton X-100 were immunoprecipitated in greater amount after extraction in other detergents. Most importantly, two antigens Mr 200 000 and 180 000 were detected only after extraction with deoxycholate or SDS.
对感染诺氏疟原虫的红细胞进行放射性碘化处理,并比较了几种非离子型、阴离子型和两性离子型去污剂提取标记膜蛋白的能力。在一些去污剂提取物中加入 Triton X-100 后,使用超免疫猴抗血清和蛋白 A-琼脂糖通过免疫沉淀法测试这些去污剂用于抗原鉴定的效果。所有测试的去污剂,包括阴离子去污剂十二烷基硫酸钠(SDS)、脱氧胆酸盐和胆酸盐;两性离子去污剂 Zwittergent-312 和 -314、CHAPS 和 Empigen BB,以及几种非离子型去污剂,都能特异性免疫沉淀 125I 标记的抗原。用不同去污剂提取后,125I 标记抗原的 SDS-聚丙烯酰胺凝胶电泳图谱各不相同,特定类型的去污剂没有一致的图谱。总共鉴定出 14 种 125I 标记的抗原,其中 11 种是使用 Triton X-100 鉴定的。一些用 Triton X-100 鉴定出的次要 125I 标记抗原,在用其他去污剂提取后免疫沉淀量更多。最重要的是,只有在用脱氧胆酸盐或 SDS 提取后才检测到两种分子量分别为 200 000 和 180 000 的抗原。