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C-peptide quantitative determination kit

A technology of quantitative determination and kit, applied in the direction of biological testing, material inspection products, etc., can solve the problems of specificity and sensitivity to be improved, poor sensitivity and anti-interference ability, and can not be used as qualitative, etc., to achieve a wide linear range, high Sensitivity and specificity, accurate and reliable results

Inactive Publication Date: 2016-06-08
GUANGZHOU KEFEN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Radioimmunoassay is limited by the working principle of the method itself, its sensitivity and anti-interference ability are poor, there are problems such as radiation radiation and pollution, and it has basically withdrawn from the market
However, enzyme-linked immunoassay is limited by methodology, and its specificity and sensitivity need to be improved. It is generally used as a preliminary screening, but it cannot be used as a qualitative or quantitative method at this stage.
When the time-resolved fluorescence immunoassay is used for ultra-micro analysis, it will be affected by the stray light of the excitation light, which will limit the sensitivity

Method used

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  • C-peptide quantitative determination kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The anti-C-peptide antibody-coated magnetic particle (M reagent) described in this example has a carboxyl group on its surface, the anti-C-peptide antibody is a monoclonal antibody, and the particle size of the magnetic particle is 1.5 μm.

[0035] The preparation method of the magnetic particle coated with the anti-C-peptide antibody is as follows:

[0036] (1) Preparation of the solution:

[0037] Coating buffer: 0.05M 2-(N-morpholine)ethanesulfonic acid (MES) buffer, wherein the pH of the coating buffer is 5.5;

[0038] Activator: add 1g ethylene dichloride (EDC) to 100mL coating buffer;

[0039] Coupling agent: Add 2g N-hydroxysuccinimide (NHS) to 100mL coating buffer;

[0040] Blocking solution: add 5g bovine serum albumin (BSA) to 100mL coating buffer;

[0041] Preservation solution: 25mM Tris-HCl solution or MES solution or 3-(N-morpholine)propanesulfonic acid solution (MOPS), 0.15M NaCl solution, 0.5% BSA by mass, 2% trehalose by mass Or sucrose, 1% sodium a...

Embodiment 2

[0062] The anti-C-peptide antibody-coated magnetic particle (M reagent) described in this example has carboxyl groups on its surface, the anti-C-peptide antibody is a monoclonal antibody, and the particle size of the magnetic particle is 3 μm.

[0063] The preparation method of the magnetic particle coated with the anti-C-peptide antibody is as follows:

[0064] (1) Preparation of the solution:

[0065] Coating buffer: 0.1M 2-(N-morpholine)ethanesulfonic acid (MES) buffer, wherein the pH of the coating buffer is 4.5;

[0066] Activator: add 2g ethylene dichloride (EDC) to 100mL coating buffer;

[0067] Coupling agent: add 1g N-hydroxysuccinimide (NHS) to 100mL coating buffer;

[0068] Blocking solution: add 10g bovine serum albumin (BSA) to 100mL coating buffer;

[0069] Preservation solution: 50mM Tris-HCl solution or MES solution or 3-(N-morpholine)propanesulfonic acid solution (MOPS), 0.2M NaCl solution, 5% BSA by mass, 0.5% trehalose by mass Or sucrose, 0.05% by mass o...

Embodiment 3

[0090] The anti-C-peptide antibody-coated magnetic particle (M reagent) described in this example has a carboxyl group on its surface, the anti-C-peptide antibody is a monoclonal antibody, and the particle size of the magnetic particle is 2 μm.

[0091] The preparation method of the magnetic particle coated with the anti-C-peptide antibody is as follows:

[0092] (1) Preparation of the solution:

[0093] Coating buffer: 0.75M 2-(N-morpholine)ethanesulfonic acid (MES) buffer, wherein the pH of the coating buffer is 4.5;

[0094] Activator: add 1.5g ethylene dichloride (EDC) to 100mL coating buffer;

[0095] Coupling agent: Add 1.5g N-hydroxysuccinimide (NHS) to 100mL coating buffer;

[0096] Blocking solution: add 8g bovine serum albumin (BSA) to 100mL coating buffer;

[0097] Preservation solution: 35mM Tris-HCl solution or MES solution or 3-(N-morpholine) propanesulfonic acid solution (MOPS), 0.18M NaCl solution, 3% BSA by mass, 1% trehalose by mass Or sucrose, 0.5% sodiu...

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Abstract

The invention relates to a C-peptide quantitative determination kit. The C-peptide quantitative determination kit comprises magnetic particles coated with anti-C-peptide antibodies, acridinium ester-marked anti-C-peptide antibodies, sample diluent, a standard C-peptide, a reaction cleaning solution, a preexciting solution and an exciting solution and is mainly used for diabetes detection. The C-peptide quantitative determination kit which is high in sensitivity and specificity, convenient to operate, wide in linearity range and quick in analysis is obtained by adopting a magnetic separation technology to be combined with an immune reaction and adopting acridinium ester or acridine sulfonamide or an acridine sulfonamide as a marker. Either serum or urine can be adopted as a detection sample, a detection result is accurate and reliable, and the C-peptide quantitative determination kit is long in preservation period and can be preserved for a year.

Description

technical field [0001] The invention belongs to the technical field of auxiliary diagnosis of diabetes, and in particular relates to a kit for quantitative determination of C-peptide in human serum or urine. Background technique [0002] Diabetes mellitus is a metabolic disease characterized by hyperglycemia due to defects in insulin secretion or insulin action. C-peptide is a secretory product of pancreatic β cells, and it has a common precursor with insulin—proinsulin. After enzymatic cleavage, one molecule of proinsulin is split into one molecule of insulin and one molecule of C-peptide. In insulin-treated diabetic patients, insulin measurement is interfered by exogenous insulin and circulating antibodies. However, insulin antibodies detected in patients on insulin therapy did not significantly interfere with C-peptide. After C-peptide is produced from the human body, it is not taken up by the liver cells, but is mainly metabolized in the kidneys and excreted from the ...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 陆建斌
Owner GUANGZHOU KEFEN BIOTECH CO LTD
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