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Exenatide detection kit

A detection kit, exenatide technology, applied in the field of biology and medical testing, can solve the problem of limiting the clinical application of GLP-1

Inactive Publication Date: 2017-01-04
ANHUI ANKE BIOTECHNOLOGY (GRP) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the GLP-1 produced by the human body is easily degraded by DPP-Ⅳ (dipeptidyl peptidase IV) in the body, and its plasma half-life is less than 2 minutes, that is, GLP-1 will be quickly inactivated by the human body, and it must be continuously administered intravenously. Injection or continuous subcutaneous injection can produce curative effect, which is inconvenient for patients with chronic diseases such as diabetes, thus greatly limiting the clinical application of GLP-1

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Examples

Experimental program
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Effect test

Embodiment 1

[0021] Example 1: Preparation of Exenatide Detection Kit

[0022] 1) Preparation of conventional equipment and reagents:

[0023] 96-well ELISA plate, exenatide positive serum, exenatide negative serum, washing solution, stop solution, horseradish peroxidase, and TMB color developing solution (which consists of color developing solution A, color developing solution B Composition), the coincidence rate of exenatide-positive sera and exenatide-negative sera was 100%.

[0024] 2) Preparation of exenatide monoclonal antibody:

[0025] a) In the first week, take 0.5ml of exenatide with a concentration of 1mg / ml and an equal volume of Freund's complete adjuvant to fully mix and emulsify, and immunize each female mouse respectively; 0.5ml of Exenatide with a concentration of 1mg / ml was fully mixed and emulsified with an equal volume of Freund's incomplete adjuvant, and each female mouse was immunized separately; blood was collected from the tail of the female mouse at the sixth week,...

Embodiment 2

[0035] Example 2: Use of Exenatide Detection Kit

[0036] 1) Add sample:

[0037] Exenatide monoclonal antibody 3D coated 7 The 96-well ELISA plate is divided into four groups of detection wells: positive control wells, negative control wells, test sample wells and blank wells. Exenatide positive serum is added to the positive control wells, and exenatide is added to the negative control wells. For negative serum, add the serum to be tested into the wells of the samples to be tested. The amount of the three samples added is 100 μl / well, then cover or cover the microplate, place the microplate at 37°C for 2 hours, discard Remove the liquid, wash with washing liquid, spin dry, and repeat the steps of washing and drying at least 3 times;

[0038] 2) Exenatide monoclonal antibody 5F labeled with horseradish peroxidase 3 :

[0039] Add 100 μl horseradish peroxidase-labeled exenatide monoclonal antibody 5F to each detection well 3 , put it at 37°C for 2 hours, discard the liqui...

Embodiment 3

[0044] Example 3: Detection of exenatide in animal serum:

[0045] Experimental grade SD rats (about 6 weeks old) were selected, and exenatide injection (trade name: Byetta) was injected subcutaneously, and blood samples were collected at 0, 5, 15, 30, 60, 120, 180, and 240 min. Then adopt the kit and ELISA method that the disclosed method of the embodiment of the present invention makes to analyze the concentration of exenatide in the blood sample, and adopt microplate reader to measure, establish metabolic curve, the result is as follows figure 2 Shown: It can be seen from the figure that after subcutaneous injection of Byetta in SD rats, the exenatide in the blood reached the peak at about 15 minutes, and reached the lower limit of detection after 3 hours. By adopting the kit involved in the present invention, the concentration of exenatide in rat blood can be effectively detected, and a complete pharmacokinetic concentration curve can be established.

[0046] It should b...

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Abstract

The invention discloses an exenatide detection kit. The exenatide detection kit comprises exenatide monoclonal antibodies, an ELISA (enzyme-linked immunosorbent assay) plate, a positive reference substance, a negative reference substance, a scrubbing solution, a stop solution and a developing solution, wherein the exenatide monoclonal antibodies comprise an exenatide monoclonal antibody coating the ELISA plate and an exenatide monoclonal antibody marked with horse radish peroxidase. When the detection kit is used for detecting concentration of exenatide of a to-be-detected sample, the sensitivity of the detection kit can be up to 1 ng / mL, the sensitivity is high, a detection result is accurate, besides, a detection object can be serum of human or animals, and an exenatide preparation is wide in application range.

Description

technical field [0001] The invention relates to the fields of biology and medical examination, in particular to an exenatide detection kit. Background technique [0002] Type 2 diabetes was originally called adult-onset diabetes, also known as non-insulin-dependent diabetes. Patients with this disease have not completely lost their ability to produce insulin, but they are in a state of relative lack of insulin in the body, and drugs can be used to stimulate insulin secretion in the body. The etiology and pathogenesis of type 2 diabetes are currently unclear, and its prominent pathophysiological feature is insulin resistance (that is, the decline in the ability of insulin to regulate glucose metabolism) accompanied by a decrease in insulin secretion or a relative decrease in islet β-cell function. [0003] Incretin is a hormone secreted by small intestinal endocrine cells in response to food intake, including glucagon-like peptide 1 (GLP-1) produced by L cells in the gastroin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68G01N2333/4609
Inventor 宋礼华王荣海杜贤宇倪晓燕程婷周乐春储成风饶海军
Owner ANHUI ANKE BIOTECHNOLOGY (GRP) CO LTD
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