Dataset Information

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

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.

SUBMITTER: Hasegawa T

PROVIDER: S-EPMC6400240 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature

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Publications

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

Hasegawa Tomonori T Komagata Miho M Hamasaki Akihiro A Harada Norio N Seino Yutaka Y Inagaki Nobuya N

Journal of diabetes investigation 20180820 2

<h4>Aims/introduction</h4>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 t ...[more]

PMID: 29993194

Similar Datasets

Project description:BackgroundThe enzyme-linked immunosorbent assay (ELISA) is an indispensable tool for clinical diagnostics to identify or differentiate diseases such as autoimmune illnesses, but also to monitor their progression or control the efficacy of drugs. One use case of ELISA is to differentiate between different states (e.g. healthy vs. diseased). Another goal is to quantitatively assess the biomarker in question, like autoantibodies. Thus, the ELISA technology is used for the discovery and verification of new autoantibodies, too. Of key interest, however, is the development of immunoassays for the sensitive and specific detection of such biomarkers at early disease stages. Therefore, users have to deal with many parameters, such as buffer systems or antigen-autoantibody interactions, to successfully establish an ELISA. Often, fine-tuning like testing of several blocking substances is performed to yield high signal-to-noise ratios.MethodsWe developed an ELISA to detect IgA and IgG autoantibodies against chitinase-3-like protein 1 (CHI3L1), a newly identified autoantigen in inflammatory bowel disease (IBD), in the serum of control and disease groups (n = 23, respectively). Microwell plates with different surface modifications (PolySorp and MaxiSorp coating) were tested to detect reproducibility problems.ResultsWe found a significant impact of the surface properties of the microwell plates. IgA antibody reactivity was significantly lower, since it was in the range of background noise, when measured on MaxiSorp coated plates (p < 0.0001). The IgG antibody reactivity did not differ on the diverse plates, but the plate surface had a significant influence on the test result (p = 0.0005).ConclusionWith this report, we want to draw readers' attention to the properties of solid phases and their effects on the detection of autoantibodies by ELISA. We want to sensitize the reader to the fact that the choice of the wrong plate can lead to a false negative test result, which in turn has serious consequences for the discovery of autoantibodies.

Dataset Information

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

Aims/introduction

Materials and methods

Results

Conclusions

Publications

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

Similar Datasets

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