A newly developed glucagon sandwich ELISA is useful for more accurate glucagon evaluation than the currently used sandwich ELISA in subjects with elevated plasma proglucagon-derived peptide levels

IF 3.2 3区医学

Journal of Diabetes Investigation Pub Date : 2023-02-02 DOI:10.1111/jdi.13986

Masaki Kobayashi, Nobuhiro Maruyama, Yukako Yamamoto, Takeshi Togawa, Takanori Ida, Morikatsu Yoshida, Mikiya Miyazato, Masahisa Kitada, Yoshitaka Hayashi, Atsunori Kashiwagi, Tadahiro Kitamura

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引用次数: 3

Abstract

Aims/Introduction

Glucagon, a peptide hormone produced from proglucagon, is involved in the pathophysiology of diabetes. Plasma glucagon levels are currently measured by sandwich enzyme-linked immunosorbent assay (ELISA), but the currently used sandwich ELISA cross-reacts with proglucagon-derived peptides, thereby providing incorrect results in subjects with elevated plasma proglucagon-derived peptide levels. We aimed to develop a more broadly reliable ELISA for measuring plasma glucagon levels.

Materials and Methods

A new sandwich ELISA was developed using newly generated monoclonal antibodies against glucagon. After its validation, plasma glucagon levels were measured with the new ELISA and the currently used ELISA in subjects who underwent laparoscopic sleeve gastrectomy (LSG) and in outpatients with suspected glucose intolerance. The ELISA results were compared with those from liquid chromatography-high resolution mass (LC-HRMS) analysis, which we previously established as the most accurate measuring system.

Results

The new ELISA has high specificity (<1% cross-reactivities) and high sensitivity (a lower range of 0.31 pmol/L). Plasma glucagon values in the subjects who underwent laparoscopic sleeve gastrectomy and some outpatients with suspected glucose intolerance differed between the new ELISA and the currently used ELISA. These subjects also showed markedly high plasma glicentin levels. Despite the elevated plasma glicentin levels, the new ELISA showed better positive correlation with LC-HRMS than did the currently used ELISA.

Conclusions

The new ELISA enables more accurate measurement of plasma glucagon than the currently used ELISA, even in subjects with elevated proglucagon-derived peptide levels. It should be clinically useful in elucidating the pathophysiology of individual diabetic patients.

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新开发的胰高血糖素夹心ELISA比目前使用的夹心ELISA在血浆胰高血糖素原衍生肽水平升高的受试者中更准确地评估胰高血糖素

目的/介绍胰高血糖素是胰高血糖素原产生的一种肽激素，参与糖尿病的病理生理。目前，血浆胰高血糖素水平是通过夹心酶联免疫吸附试验(ELISA)测量的，但目前使用的夹心酶联免疫吸附试验与胰高血糖素衍生肽交叉反应，因此在血浆胰高血糖素衍生肽水平升高的受试者中提供不正确的结果。我们的目标是开发一种更广泛可靠的ELISA来测量血浆胰高血糖素水平。材料与方法利用新制备的抗胰高血糖素单克隆抗体，建立了一种新的夹心酶联免疫吸附试验。在验证后，用新的ELISA和目前使用的ELISA测定了接受腹腔镜袖胃切除术(LSG)的受试者和疑似葡萄糖耐受不良的门诊患者的血浆胰高血糖素水平。将ELISA结果与液相色谱-高分辨率质谱(LC-HRMS)分析结果进行比较，液相色谱-高分辨率质谱分析是我们之前建立的最精确的测量系统。结果新酶联免疫吸附试验具有高特异性(交叉反应率为1%)和高灵敏度(低范围为0.31 pmol/L)。在接受腹腔镜胃切除术的受试者和一些疑似葡萄糖耐受不良的门诊患者中，新的ELISA和目前使用的ELISA的血浆胰高血糖素值不同。这些受试者的血浆甘肽水平也明显升高。尽管血浆glicentin水平升高，但与目前使用的ELISA相比，新ELISA与LC-HRMS呈正相关。结论:与目前使用的ELISA相比，新的ELISA能够更准确地测量血浆胰高血糖素，即使在胰高血糖素原衍生肽水平升高的受试者中也是如此。这对阐明糖尿病患者个体的病理生理应具有临床意义。

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来源期刊

Journal of Diabetes Investigation Medicine-Internal Medicine

自引率

9.40%

发文量

218

期刊介绍： Journal of Diabetes Investigation is your core diabetes journal from Asia; the official journal of the Asian Association for the Study of Diabetes (AASD). The journal publishes original research, country reports, commentaries, reviews, mini-reviews, case reports, letters, as well as editorials and news. Embracing clinical and experimental research in diabetes and related areas, the Journal of Diabetes Investigation includes aspects of prevention, treatment, as well as molecular aspects and pathophysiology. Translational research focused on the exchange of ideas between clinicians and researchers is also welcome. Journal of Diabetes Investigation is indexed by Science Citation Index Expanded (SCIE).