Establishment of a novel 70K Mac-2 binding protein antibody through screening of fucosylation-related antibodies

岩藻糖基化 抗体 单克隆抗体 糖基化 聚糖 糖蛋白 癌症研究 分子生物学 抗体微阵列 免疫细胞化学 免疫沉淀 生物 化学 生物化学 免疫学 内分泌学
作者
Mika Masuda,Tatsuya Asuka,Naoko Terao,Shinsuke Nishino,Shoichiro Ikeda,Shinji Takamatsu,Jumpei Kondo,Eiji Miyoshi
出处
期刊:Journal of Biochemistry [Oxford University Press]
卷期号:173 (6): 487-495
标识
DOI:10.1093/jb/mvad015
摘要

Abstract Mac-2 binding protein (Mac-2bp) is a serum glycoprotein that contains seven N-glycans, and Mac-2bp serum levels are increased in patients with several types of cancer and liver disease. Mac-2bp glycosylation isomer has been applied as a clinical biomarker of several diseases, including liver fibrosis. In the present study, we identified fucosylated Mac-2bp in the conditioned medium of cancer cells resistant to anticancer therapies using glycoproteomic analyses. Fucosylation is one of the most important types of glycosylation involved in carcinogenesis and cancer stemness. To establish a next-generation glycan antibody for fucosylated Mac-2bp, we used fucosylation-deficient HEK293T cells to prepare reference Mac-2bp antigens and performed antibody screening. Unexpectedly, the 19-8H mAb obtained with our screen recognized 70K Mac-2bp, which is C-terminus-truncated product, rather than specifically recognizing fucosylated Mac-2bp. We performed immunocytochemistry using our novel 19-8H mAb, which resulted in strong cell surface staining of anticancer drug-resistant cancer cells. Therefore, our novel 19-8H mAb represents a valuable tool for cancer biology research that can help elucidate the biological function of 70K Mac-2bp.
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