Fukaya Shinichi, Yabe Tomio, Kanamaru Yoshihiro
United Graduate School of Agricultural Science, Gifu University, Gifu, Japan.
Biosci Biotechnol Biochem. 2010;74(10):2141-4. doi: 10.1271/bbb.100436. Epub 2010 Oct 7.
We have recently demonstrated that the 1CF11 monoclonal antibody bound human milk lactoferrin (hLf) through the recognition of two distinct portions of the molecule, namely the N-glycan-relevant and -irrelevant structural elements. In this present study, we prepared four immunoreactive peptide fractions containing N-linked glycan from tryptic digests of reduced and alkylated hLf by using a concanavalin A lectin column and reverse-phase HPLC. Deglycosylation of these fractions and a competitive binding assay using fucosylated oligosaccharides revealed that the non-reducing terminal fucose residue in N-linked glycan(s) played a significant role in recognizing the N-glycan-relevant element in hLf by 1CF11.
我们最近证明，1CF11单克隆抗体通过识别分子的两个不同部分，即与N-聚糖相关和不相关的结构元件，与人类母乳乳铁蛋白（hLf）结合。在本研究中，我们使用伴刀豆球蛋白A凝集素柱和反相高效液相色谱法，从还原和烷基化的hLf的胰蛋白酶消化物中制备了四个含有N-连接聚糖的免疫反应性肽级分。这些级分的去糖基化以及使用岩藻糖基化寡糖的竞争性结合试验表明，N-连接聚糖中的非还原末端岩藻糖残基在1CF11识别hLf中与N-聚糖相关的元件方面发挥了重要作用。
