Adachi Y, Ohno N, Yadomae T
Laboratory of Immunopharmacology of Microbial Products, Tokyo College of Pharmacy, Japan.
Biol Pharm Bull. 1994 Nov;17(11):1508-12. doi: 10.1248/bpb.17.1508.
Antibody for (1-->6)-branched (1-->3)-beta-D-glucan was prepared using rodents. An antitumor (1-->6)-beta-monoglucosyl branched (1-->3)-beta-D-glucan (GRN: grifolan) was conjugated with bovine serum albumin and used as an immunogen. The antibody titer in serum was determined by ELISA using biotin-conjugated GRN. Administration of the antigen raised the antibody titer only in the rabbit, with mouse and rat showing no significant antibody titer for the glucan. The antigen specificity of the anti-GRN antibody was determined by competitive ELISA. The rabbit anti-GRN antibody bound to structurally related antitumor (1-->6)-branched (1-->3)-beta-D-glucans such as lentinan, schizophyllan and SSG, whereas it did not react with linear (1-->3)-beta-D-glucan, curdlan or GRN-derivatives obtained by periodate-oxidation and Smith degradation. These facts strongly suggest that the hapten site of the antibody was the monoglucosyl branched moiety of (1-->3)-beta-D-glucan. These results indicate that this antibody would be a useful probe for the detection of (1-->6)-branched antitumor glucans administered to the host.
使用啮齿动物制备针对(1→6)分支的(1→3)-β-D-葡聚糖的抗体。将一种抗肿瘤的(1→6)-β-单葡萄糖基分支的(1→3)-β-D-葡聚糖(GRN:灰树花多糖)与牛血清白蛋白偶联,并用作免疫原。血清中的抗体效价通过使用生物素偶联的GRN的ELISA法测定。给予抗原仅在兔中提高了抗体效价,而小鼠和大鼠对该葡聚糖未显示出明显的抗体效价。抗GRN抗体的抗原特异性通过竞争性ELISA法测定。兔抗GRN抗体与结构相关的抗肿瘤(1→6)分支的(1→3)-β-D-葡聚糖如香菇多糖、裂褶菌多糖和SSG结合,而它不与线性(1→3)-β-D-葡聚糖、凝胶多糖或通过高碘酸盐氧化和史密斯降解获得的GRN衍生物反应。这些事实强烈表明该抗体的半抗原位点是(1→3)-β-D-葡聚糖的单葡萄糖基分支部分。这些结果表明该抗体将是用于检测给予宿主的(1→6)分支的抗肿瘤葡聚糖的有用探针。