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Development of a Fucoidan-Specific Antibody and Measurement of Fucoidan in Serum and Urine by Sandwich ELISA

机译:夹心酶联免疫吸附测定法制备岩藻依聚糖特异性抗体并测定血清和尿液中岩藻依聚糖的含量

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摘要

Fucoidan exhibits various biological properties. We raised a novel antibody against fucoidan extracted from Cladosiphon okamuranus and developed a sandwich ELISA method to measure fucoidan. The fucoidan antibody was specific and did not cross-react with other polysulfated polysaccharides. Fucoidan recovery from serum and urine by ELISA was 86-113%. Intra- and inter-assay CVs were 1.5-13.4%. Assay linearity was maintained after 3-fold dilution of each sample with phosphate-buffer saline (PBS). In the serum and urine of healthy volunteers (n = 10), fucoidan was not detected before administration, and the levels markedly increased 6 and 9h after oral administration. The molecular weight of the serum fucoidan determined by HPLC gel filtration remained unchanged, whereas that of urine fucoidan was significantly reduced. This is the first ELISA method of measuring serum and urine fucoidan levels after oral administration. The method is simple, reliable, and practical for the analysis of samples, especially urine samples.
机译:岩藻依聚糖具有多种生物学特性。我们提出了一种新的抗从岩藻中提取的岩藻依聚糖的抗体,并开发了一种夹心ELISA方法来测量岩藻依聚糖。岩藻依聚糖抗体具有特异性,不会与其他多硫酸化多糖交叉反应。通过ELISA从血清和尿液中的岩藻依丹回收率为86-113%。批内和批间CV为1.5-13.4%。每个样品用磷酸盐缓冲盐水(PBS)稀释3倍后,测定线性得以保持。在健康志愿者的血清和尿液中(n = 10),在给药前未检测到岩藻依聚糖,并且口服后6h和9h含量明显升高。通过HPLC凝胶过滤测定的血清岩藻依聚糖的分子量保持不变,而尿岩藻依聚糖的分子量显着降低。这是口服后测量血清和尿中岩藻依聚糖水平的第一种ELISA方法。该方法简单,可靠,对样品特别是尿液样品的分析实用。

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