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Detection kit for completely and homogeneously determining glucagon and method thereof

A glucagon and detection kit technology, which is applied in the field of detection kits for completely homogeneous determination of glucagon, can solve the problems of inability to meet detection requirements, limited development and application, and low degree of automation, and achieves rapid detection. Convenience, improved accuracy and sensitivity, wide inspection results

Inactive Publication Date: 2019-04-19
GUANGZHOU JINDE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Both of the above two methods have certain defects: poor accuracy, long operation time, low degree of automation, unable to meet the clinical needs of rapid detection of a large number of samples
There are many factors affecting the detection process of this method, but the most important is the existence of radioactive pollution, which is not easy to solve, thus limiting its development and application

Method used

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  • Detection kit for completely and homogeneously determining glucagon and method thereof
  • Detection kit for completely and homogeneously determining glucagon and method thereof
  • Detection kit for completely and homogeneously determining glucagon and method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Embodiment 1, the preparation of glucagon calibrator

[0064] The matrix liquid of calibrator of the present invention comprises the component of following concentration:

[0065]

[0066]

[0067] Prepare the calibrator matrix solution according to the above ratio, and use this matrix solution to dilute the glucagon antigen into corresponding concentrations of calibrator, the concentrations are 1pmol / L, 4pmol / L, 16pmol / L, 32pmol / L, 128pmol / L, Add 0 pmol / L matrix solution, a total of 6 bottles, 1 mL each, and store at 2-8°C.

Embodiment 2

[0068] Example 2, Preparation of Acridan-labeled anti-glucagon antibody

[0069] Take 250μg of anti-glucagon antibody (diluted to 1mg / mL with 20mM PBS first), add 40μL of Acridan, add 710μL of 20mM PBS, and incubate on a rotator at room temperature for 1h; then use marker buffer (30mM pH 7.6 PBS buffer, 0.1w / v% sodium thiosulfate, 0.5w / v% BSA, 0.4w / v% mannitol and 0.05w / v% ProClin300) were diluted to the working concentration.

[0070] According to this method, cridan-labeled anti-glucagon antibodies with concentrations of 1.5 mg / L, 3 mg / L, 6 mg / L, 12 mg / L, 24 mg / L and 48 mg / L were prepared.

Embodiment 3

[0071] Example 3, preparation of horseradish peroxidase-labeled anti-glucagon antibody

[0072] Use a commercial HRP labeling kit to label anti-glucagon antibody, and then use label buffer (30mM pH7.6PBS buffer, 0.05w / v% VC or 0.05w / v% reduced glutathione, 0.03w / v% EC Oxyrase, 0.5w / v% mannitol, 0.5w / v% polyethylene glycol 20000, 0.1w / v% enzyme stabilizer, 0.05w / v% ProClin 300.) diluted to the working concentration.

[0073] According to this method, HRP-labeled anti-glucagon antibodies were prepared at concentrations of 0.025 mg / L, 0.05 mg / L, 0.1 mg / L, 0.2 mg / L, 0.4 mg / L, and 0.8 mg / L, respectively.

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Abstract

The invention belongs to the field of biotechnologies and discloses a detection kit for completely and homogeneously determining glucagon and a method thereof. The detection kit utilizes a spatial neighborhood chemiluminescence technology to achieve complete and homogeneous detection of glucagon and is based on a double-sandwich technique, two monoclonal antibodies are bound to antigenic determinants of glucagon molecules respectively, a calibrator or a sample to be tested, a horse radish peroxidase-labeled anti-human glucagon antibody, a 9,10-dihydroacridine-labeled anti-human glucagon antibody are added together into a microplate, the two monoclonal antibodies and the antigen in the sample form an antibody-antigen-antibody sandwich complex without coating after incubation, the washing process is omitted, a trigger is added after an antioxidant is added and evenly mixed, a luminescence intensity value in a period of time is immediately continuously detected, and the content of the glucagon in the sample to be tested is calculated. The detection kit has the advantages of high stability, rapid and convenient detection, high sensitivity and strong specificity.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a detection kit for completely homogeneously measuring glucagon and a method thereof. Background technique [0002] Glucagon (Glu) is also known as glucagon, anti-insulin or insulin B. It is a hormone secreted by the alpha cells of the vertebrate pancreas along with insulin. It acts against insulin and acts to increase blood sugar. In 1953, it was separated and precipitated to obtain crystallization. It is a single-chain peptide (molecular weight of about 3500) consisting of 29 amino acid residues starting from N-terminal histidine and ending at C-terminal threonine. There is no S-S bond in the molecule. At this point, Completely different from insulin. The structure of this compound has been confirmed by recent chemical synthesis. The initial process of glucagon's action is to specifically bind to receptors present on the target cell membrane to activate adenylyl cyclase, and c...

Claims

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Application Information

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IPC IPC(8): G01N33/74G01N33/532G01N33/535G01N21/76
CPCG01N21/76G01N33/532G01N33/535G01N33/74
Inventor 蓝媛李民友段朝晖张玲蔺涛梁灿高国全
Owner GUANGZHOU JINDE BIOTECH
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