Arnold F, Bédouet L, Batina P, Robreau G, Talbot F, Lécher P, Malcoste R
Université de Bretagne Occidentale, Institut Universitaire de Technologie, Laboratoire Universitaire de Microbiologie Appliquée de Quimper, France.
Microbiol Immunol. 1998;42(1):23-31. doi: 10.1111/j.1348-0421.1998.tb01965.x.
The monoclonal antibody 21E7-B12 (IgG3) can be used in a direct method of Clostridium tyrobutyricum detection based on an immunoenzymatic assay. Immunoelectron microscopy demonstrated that the 21E7-B12 antibody recognized the surface-exposed epitopes on the flagellar filaments of C. tyrobutyricum. After flagellar extraction, the purified flagellin showed an apparent molecular mass of 46 kDa with an isoelectric point of 3.6. Sugar staining, mild periodate oxidation and beta-elimination experiments showed that the flagellin was glycosylated and that the 21E7-B12 epitope was located in the sugar moiety. Amino acid composition showed that the flagellar filament protein contained a high percentage of serine and threonine, while proline was absent. The first 23 residues of the N-terminal were determined and sequence homology with other flagellins was found.
单克隆抗体21E7-B12（IgG3）可用于基于免疫酶测定的酪丁酸梭菌直接检测方法。免疫电子显微镜显示，21E7-B12抗体识别酪丁酸梭菌鞭毛丝上暴露于表面的表位。鞭毛提取后，纯化的鞭毛蛋白显示表观分子量为46 kDa，等电点为3.6。糖染色、温和的高碘酸盐氧化和β-消除实验表明，鞭毛蛋白是糖基化的，且21E7-B12表位位于糖部分。氨基酸组成表明，鞭毛丝蛋白含有高比例的丝氨酸和苏氨酸，而不含脯氨酸。确定了N端的前23个残基，并发现了与其他鞭毛蛋白的序列同源性。
