The effect of fish matrix on the enzyme-linked immunosorbent assay of antibiotics.
The effect of fish matrix on the enzyme-linked immunosorbent assay of antibiotics.
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DOI：
10.1002/jsfa.5931
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发表时间：
2013-05
影响因子：
4.1
通讯作者：
Xiudan Wang;Hong Lin;Jianxin Sui;Limin Cao
中科院分区：
文献类型：
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作者：
Xiudan Wang;Hong Lin;Jianxin Sui;Limin Cao
背景基质效应被认为是食品免疫测定中的一个问题。然而，有关水产品的干扰及其机制的信息非常有限。以大菱鲆（Scophthalmus maximus）、牙鲆（Paralichthys olivaceus）和粗壮舌鳎（Cymoglossus robustus）3种比目鱼为样品，研究了鱼基质对抗生素（诺氟沙星）残留竞争性间接酶联免疫吸附测定（ci-ELISA）的影响。并对基体效应的机理进行了初步探讨。结果在校准曲线的工作范围内，在鱼提取物存在下，观察到抑制率的显著（P = 0.05）但不规则的变化。进一步的实验表明，这种基质效应可能是由一些具有宽范围分子量（从低于14.4 kDa到约116.0 kDa）的水溶性鱼蛋白引起的，并且来自鱼肌肉的离子也可能有助于干扰。免疫印迹结果表明，某些鱼蛋白组分可与所用的抗体试剂有效结合。结论鱼基质对诺氟沙星免疫分析有明显干扰。一些蛋白质和离子被证明有助于研究的基质效应。虽然详细的机制尚不清楚，但鱼蛋白与免疫球蛋白G（IgG）或辣根过氧化物酶（HRP）标记的IgG之间的非特异性相互作用被认为是免疫测定中基质效应的重要来源。
BACKGROUND The matrix effect is considered to be a problem in the immunoassay of foodstuffs. However, information on the interference from aquatic products, as well as the mechanism involved, is very limited. In this study, using three flatfishes (Scophthalmus maximus, Paralichthys olivaceus and Cymoglossus robustus) as samples, the effect of the fish matrix on the competitive indirect enzyme-linked immunosorbent assay (ci-ELISA) of antibiotic (norfloxacin) residues was investigated. The mechanism of the observed matrix effect is also preliminarily discussed. RESULTS Within the working range of the calibration curves, a significant (P = 0.05) but irregular variation in the inhibition ratio was observed in the presence of fish extracts. Further experiments revealed that such a matrix effect could be caused by some water-soluble fish proteins with a wide range of molecular weight (from below 14.4 kDa to about 116.0 kDa), and the ions from fish muscles may also contribute to the interference. The results of western blotting indicated that some fish protein components might effectively bind with antibody reagents used. CONCLUSION Significant interference in the immunoassay of norfloxacin was observed in the presence of fish matrix. Some proteins and ions were demonstrated to contribute to the matrix effect investigated. Although the detailed mechanism is still unclear, the non-specific interaction between fish proteins and immunoglobulin G (IgG) or horseradish peroxidase (HRP) labelled IgG was assumed to be an important source of the matrix effect in immunoassays.