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固相萃取处理是必需的,以测量受异嗜性抗体影响的酶联免疫吸附测定试剂盒中的活性胰高血糖素样肽-1。

Solid-phase extraction treatment is required for measurement of active glucagon-like peptide-1 by enzyme-linked immunosorbent assay kit affected by heterophilic antibodies.

机构信息

SRL Inc., Tokyo, Japan.

Department of Diabetes, Endocrinology and Nutrition, Graduate School of Medicine, Kyoto University, Kyoto, Japan.

出版信息

J Diabetes Investig. 2019 Mar;10(2):302-308. doi: 10.1111/jdi.12896. Epub 2018 Aug 20.

Abstract

AIMS/INTRODUCTION: It is reported that interfering substances in the blood might influence the value for measurement of active glucagon-like peptide-1 (GLP-1) in human plasma. Solid phase extraction (SPE) pretreatment is recommended to reduce their influence, but it requires a lot of cost and time. However, there is little investigation about causative inhibitory substances and about methods that can replace solid phase extraction. In the present study, we aimed to seek the candidate of the substances that might interfere with an active GLP-1 enzyme-linked immunosorbent assay (ELISA).

MATERIALS AND METHODS

Two kinds of active GLP-1 ELISA kits using different antibodies, plural extraction carriers and elution solutions were used to evaluate the SPE method. Active GLP-1 concentration was compared with or without SPE, and with or without a heterophilic blocking tube.

RESULTS

Active GLP-1 values were often higher without SPE compared with those with SPE pretreatment. This difference was eliminated by pretreatment with a heterophilic blocking tube or ELISA kits that did not use a mouse monoclonal antibody, and was independent of SPE.

CONCLUSIONS

Substances absorbed to a heterophilic blocking tube carrier might interfere with an active GLP-1 immunoassay. Solid-phase extraction treatment is required for measurement of active GLP-1 by an ELISA kit affected by heterophilic antibodies.

摘要

目的/引言:据报道,血液中的干扰物质可能会影响人血浆中活性胰高血糖素样肽-1(GLP-1)的测量值。推荐使用固相萃取(SPE)预处理来减少其影响,但这需要大量的成本和时间。然而,关于导致抑制作用的物质以及可以替代固相萃取的方法的研究很少。在本研究中,我们旨在寻找可能干扰活性 GLP-1 酶联免疫吸附测定(ELISA)的物质的候选物。

材料和方法

使用两种不同抗体、多种提取载体和洗脱液的两种活性 GLP-1 ELISA 试剂盒来评估 SPE 方法。比较了有无 SPE 预处理以及有无异嗜性封闭管时的活性 GLP-1 浓度。

结果

与 SPE 预处理相比,无 SPE 时活性 GLP-1 值通常更高。通过使用异嗜性封闭管或不使用小鼠单克隆抗体的 ELISA 试剂盒进行预处理,可以消除这种差异,并且这种差异与 SPE 无关。

结论

吸附到异嗜性封闭管载体上的物质可能会干扰活性 GLP-1 免疫测定。受异嗜性抗体影响的活性 GLP-1 的测定需要固相萃取处理。

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