Reagent, method and kit for detection of biological activity of glucagon-like peptide-1 (GLP-1)

A technology of glucagon and biological activity, applied in the field of detection reagents, can solve problems such as short retention time, and achieve the effects of preventing the loss of cAMP, improving sensitivity and strong practicability

Active Publication Date: 2013-10-09
MEIDE TAIPINGYANG PRECISION INSTR MFG
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

Aiming at the limitation that GLP-1 analogues stay in the body for a short time in clinical practice and require daily injections, a GLP-1 mutant polypeptide with a longer half-life

Method used

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  • Reagent, method and kit for detection of biological activity of glucagon-like peptide-1 (GLP-1)
  • Reagent, method and kit for detection of biological activity of glucagon-like peptide-1 (GLP-1)
  • Reagent, method and kit for detection of biological activity of glucagon-like peptide-1 (GLP-1)

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Embodiment Construction

[0059] The present invention will be further described in detail below through the specific examples, the following examples are only descriptive, not restrictive, and cannot limit the protection scope of the present invention with this.

[0060] The invention establishes a method for measuring GLP-1 biological activity based on receptor-ligand binding, and directly captures cAMP secreted after receptor activation, and corresponding detection reagents. The present invention clones the target gene of GLP-1 receptor, and establishes a cell line expressing GLP-1 receptor through gene recombination technology, thereby obtaining purified GLP-1 receptor, GLP-1 and GLP-1 in the serum (or plasma) to be tested The GLP-1 receptor on the cell membrane binds, the receptor is activated to produce the second messenger cAMP, the microwell plate coated with goat anti-human cAMP polyclonal antibody directly captures the cAMP produced, and then adds biotin-labeled mouse anti-human The cAMP mono...

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Abstract

The invention relates to a reagent for detection of biological activity of glucagon-like peptide-1 (GLP-1). The reagent added into each 15 microliters of a detection sample comprises: 1, 35-45 microliters of a membrane reaction solution, 2, 15-25 microliters of a GLP-1 acceptor, 3, 10-20 microliters of a GLP-1 standard substance, 4, 5-15 microliters of sitgliptin phosphate, 5, 1-10 microliters of triphosadenine, 6, 1-10 microliters of guanosine triphosphate, 7, 5-15 microliters of 3-isobutyl-1-methylxanthine, 8, 1-10 microliters of bovine serum albumin, 9, 50-150 microliters of a biotin-labeled mouse anti-human cAMP monoclonal antibody, 10, a microwell plate coated with a goat anti-human cAMP polyclonal antibody, 11, 50-150 microliters of an avidin-labeled horseradish peroxidase, 12, 300-400 microliters of a PBS washing liquor, 13, 50-150 microliters of a chromogenic substrate, and 14, 50-150 microliters of a stop buffer. The GLP-1 acceptor is produced by genetic recombination, is convenient for purification and can reduce the batch difference of the reagent. The GLP-1 acceptor is used as a probe for capturing GLP-1 so that detection sensitivity is improved, biological activity can be directly determined, sensitivity is high, and practicality is strong.

Description

technical field [0001] The invention relates to a detection reagent in the field of medical testing, in particular to a detection reagent, detection method and kit for glucagon-like peptide-1 (glucagon-like peptide-1, GLP-1) biological activity. Background technique [0002] Diabetes is based on polygenic inheritance, coupled with the effects of environmental factors and autoimmunity, through an incompletely elucidated mechanism, causing insulin secretion disorders and insufficient insulin biological effects, resulting in hyperglycemia as the basic biochemical characteristics of sugar, A group of clinical syndromes characterized by disorders of fat, protein, water and electrolyte metabolism. In typical cases, symptoms of "three excesses and one deficiency" such as polyuria, polydipsia, polyphagia, and weight loss may appear. The chronic complications are mainly specific and non-specific microangiopathy and peripheral neuropathy. At present, diabetes has become the third maj...

Claims

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Application Information

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IPC IPC(8): G01N33/68
Inventor 李迺昶李会强周宇捷任峰
Owner MEIDE TAIPINGYANG PRECISION INSTR MFG
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