Wold J K, Slayter H S, Codington J F, Jeanloz R W
Biochem J. 1985 Apr 1;227(1):231-7. doi: 10.1042/bj2270231.
Antibodies of the IgM type present in rabbit anti-epiglycanin antiserum were purified by (NH4)2SO4 precipitation and by ion-exchange, affinity and gel-filtration chromatography. After papain treatment of this fraction, followed by gel filtration, the fraction with highest apparent Mr was incubated with epiglycanin, and the antigen-antibody complexes separated by gel filtration. These were examined by electron microscopy, using rotational shadow casting, and the photographs of the complexes were mapped for the locations of the antibody molecules on the extended epiglycanin molecules. Distribution of the frequency of attachment of immunoglobulin showed a strong tendency toward binding at one end of epiglycanin, suggesting the probable presence of only one epitope, probably a glycopeptide structure containing a beta-D-galactopyranosyl-(1----3)-2-acetamido-2-deoxy-D-galactose chain.
通过硫酸铵沉淀以及离子交换、亲和和凝胶过滤色谱法，对兔抗表皮糖蛋白抗血清中存在的IgM型抗体进行了纯化。用木瓜蛋白酶处理该组分后，接着进行凝胶过滤，将表观分子量最高的组分与表皮糖蛋白一起孵育，然后通过凝胶过滤分离抗原-抗体复合物。使用旋转阴影投射通过电子显微镜对这些复合物进行检查，并绘制复合物的照片以确定抗体分子在伸展的表皮糖蛋白分子上的位置。免疫球蛋白附着频率的分布显示出在表皮糖蛋白一端结合的强烈趋势，这表明可能仅存在一个表位，可能是一个含有β-D-吡喃半乳糖基-(1→3)-2-乙酰氨基-2-脱氧-D-半乳糖链的糖肽结构。
