Development of a Fucoidan-Specific Antibody and Measurement of Fucoidan in Serum and Urine by Sandwich ELISA
Development of a Fucoidan-Specific Antibody and Measurement of Fucoidan in Serum and Urine by Sandwich ELISA
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DOI：
10.1271/bbb.90705
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发表时间：
2010-02-01
影响因子：
1.6
通讯作者：
Nagamine, Takeaki
中科院分区：
文献类型：
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作者：
Tokita, Yoshiharu;Nakajima, Katsuyuki;Nagamine, Takeaki
褐藻糖胶具有多种生物学特性。我们提出了一种从冈村枝管藻中提取的新型抗岩藻依聚糖抗体，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，不与其他多硫酸化多糖发生交叉反应。通过 ELISA 从血清和尿液中回收岩藻依聚糖为 86-113%。测定内和测定间 CV 为 1.5-13.4%。用磷酸盐缓冲盐水 (PBS) 将每个样品稀释 3 倍后，仍保持测定线性。在健康志愿者（n = 10）的血清和尿液中，给药前未检测到岩藻依聚糖，口服后6和9小时水平显着升高。 HPLC凝胶过滤测定血清岩藻依聚糖的分子量保持不变，而尿液岩藻依聚糖的分子量显着降低。这是第一个测定口服后血清和尿液岩藻依聚糖水平的 ELISA 方法。该方法简单、可靠、实用，适用于样品特别是尿液样品的分析。
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 9 h 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.