Kanamaru Y, Ikeda S, Yang S M, Kaneko T, Kuwata T, Nagaoka S, Shimizu M, Sachdev G P
Department of Food Science, Faculty of Agriculture, Gifu University, Japan.
Biochem Biophys Res Commun. 1998 Aug 28;249(3):618-23. doi: 10.1006/bbrc.1998.9208.
A monoclonal antibody (mAb; a mouse IgM referred to as 1CF11) recognizing various human glycoproteins was obtained. While the immunoreaction of glycoproteins from human secretions including milk, saliva, and bronchus was demonstrated as a typical dose-responded S-shaped reaction curve on ELISA, no reaction was detected with milks and sera of animal origin as well as human serum. In the constituting polypeptides of the human milk secretory IgA molecule, only the secretory component was recognized by this mAb. Among various chemical treatments of the purified human milk lactoferrin (Lf), only either periodate or mild alkaline treatment abolished the immunoreactivity of the glycoprotein. A recombinant human Lf was not immunoreactive. Finally, the immunoreactive fragments were isolated from human milk Lf, which remained reactive with PAS reagent while lacking the previously reported N-glycans. These results strongly suggest that the mAb 1CF11 recognizes a new glycan O-glycosidically linked to glycoproteins in human secretions.
获得了一种识别多种人类糖蛋白的单克隆抗体(mAb;一种称为1CF11的小鼠IgM)。虽然在酶联免疫吸附测定(ELISA)中,来自包括牛奶、唾液和支气管分泌物等人类分泌物中的糖蛋白的免疫反应表现为典型的剂量反应S形反应曲线,但未检测到与动物来源的牛奶和血清以及人类血清发生反应。在人乳分泌型IgA分子的组成多肽中,只有分泌成分被该单克隆抗体识别。在对纯化的人乳铁蛋白(Lf)进行的各种化学处理中,只有高碘酸盐处理或温和的碱性处理消除了该糖蛋白的免疫反应性。重组人Lf无免疫反应性。最后,从人乳Lf中分离出免疫反应性片段,这些片段与过碘酸希夫(PAS)试剂仍有反应,但缺乏先前报道的N-聚糖。这些结果有力地表明,单克隆抗体1CF11识别一种与人分泌物中糖蛋白以O-糖苷键连接的新聚糖。