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首页> 外文期刊>The Open Clinical Chemistry Journal >Inhibition of Horseradish Peroxidase (HRP) by a Nonhydrophobic Component of Urine: A Caution for Immunoassays
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Inhibition of Horseradish Peroxidase (HRP) by a Nonhydrophobic Component of Urine: A Caution for Immunoassays

机译：尿液中非疏水成分对辣根过氧化物酶（HRP）的抑制作用：免疫测定的注意事项

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Undiluted urine mixed directly with HRP suppresses generation of enzyme product to less than 80% of that seen in buffer controls. Incubating dilutions of various urine preparations with HRP immobilized on concanavalin A coated microtiter plates reveals that the source of urine or HRP, and the type of HRP substrate used have minimal effect on the degree of HRP suppression; only dilutions of urine greater than 8-fold with buffer produce HRP activities equivalent to those in buffer. Treating urine with charcoal or C18 silica only partially reverses the HRP suppression. Inhibition of HRP in competitive assays biases results to the high side and in noncompetive assays biases results to the low side. The present findings suggest analysts should avoid immunoassay protocols that allow direct contact between undiluted urine and HRP reporter conjugates and should be cautious with quantitative results previously reported from assays that used undiluted urine with HRP reporters.
机译：直接与HRP混合的未稀释尿液将酶产物的产生抑制到缓冲液对照中产生的酶产物的80％以下。将各种尿液制剂的稀释液与固定在伴刀豆球蛋白上的HRP孵育后，可发现尿液或HRP的来源以及所用HRP底物的类型对HRP抑制程度的影响最小。只有用缓冲液稀释的尿液大于8倍才能产生与缓冲液相同的HRP活性。用木炭或C18二氧化硅处理尿液只能部分逆转HRP抑制作用。在竞争性测定中抑制HRP会使结果偏向高端，而在非竞争性测定中，偏向结果偏低。本研究结果表明，分析人员应避免允许未稀释尿液与HRP报告分子偶联物直接接触的免疫测定方案，并应谨慎对待先前从使用HRP报告分子使用未稀释尿液的测定中报告的定量结果。

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