Competitive enzyme-linked immunosorbent assay for the determination of catecholamine, dopamine in serum
Competitive enzyme-linked immunosorbent assay for the determination of catecholamine, dopamine in serum
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DOI：
10.1016/j.aca.2008.02.042
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发表时间：
2008-06-30
影响因子：
6.2
通讯作者：
Paeng, Insook Rhee
中科院分区：
文献类型：
--
作者：
Kim, Jisun;Jeon, Moonsun;Paeng, Insook Rhee
多巴胺（DA）的竞争性酶联免疫吸附测定已被优化和表征。DA是敏感的氧气和光根据DA氧化过程中的pH值的函数。DA中的酚基容易氧化成醌型，因此游离DA在碱性介质中劣化。因此，考虑了诸如pH、酶标底物、离子强度和反应时间等因素对ELISA性能的影响。用5 μ g/mL（-1）BSA-DA和1/7500稀释的抗DA抗体进行测定。绘制了剂量-反应曲线，DA的检测限和动态范围分别达到1.0 × 10（-9）M（0.19 μ g L-1）和5个数量级（3.2 × 10（-8）M至3.2 × 10（-3）M）。在缓冲液和血清中获得的吸光度的相关图显示出良好的一致性，相关系数（R-2 =0.9947）：Abs. (in血清）= 0.6128 x Abs. (in缓冲液）+0.2926。用结构相似的化合物检查交叉反应性。结果表明，肾上腺素与3-甲氧基酪胺的交叉反应率为18.9%，而3，4-二羟基苯乙酸与高香草酸的交叉反应率较低（
A competitive enzyme-linked immunosorbent assay for dopamine (DA) has been optimized and characterized. DA is sensitive to oxygen and light according to a function of the pH on the DA oxidation process. The phenolic groups in DA are readily oxidisable to a quinoid form and thus, free DA deteriorates in alkaline media. Thus, effect of factors such as pH, enzyme-label with substrate, ionic strength and reaction time was considered on performance of ELISA. Assay was performed with 5 mu g mL(-1) of BSA-DA and 1/7500 dilution of anti-DA antibody. A dose-response curve was constructed, and a limit of detection and a dynamic range for DA were accomplished to 1.0 x 10(-9) M (0.19 mu g L-1) and five orders (3.2 x 10(-8) M to 3.2 x 10(-3) M) of magnitude, respectively. The correlation diagram of the absorbance obtained both in buffer and in serum has shown good agreement with correlation coefficient (R-2 =0.9947): Abs. (in serum) = 0.6128 x Abs. (in buffer) + 0.2926. The cross-reactivity was examined with the structurally similar compounds. And the results demonstrated that epinephrine and 3-methoxytyramine showed cross-reactivity (18.9% each), whereas 3,4-dihydroxyphenylacetic acid and homovanillic acid showed low cross-reactivity (