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C-peptide micropore plate type magnetic granule chemoluminescence immunoassay measuring kit and preparation method thereof

A chemiluminescent immunoassay kit technology, applied in the field of immunoanalysis medicine, can solve the problems of radioactive pollution in the environment, easy to be interfered by air dust, narrow linear range, etc., and achieve no radioactive pollution, strong clinical applicability, and detection range wide effect

Inactive Publication Date: 2009-09-30
北京科美东雅生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Because radioimmunoassay technology uses radioactive elements as markers, it has radioactive pollution to the environment, and has disadvantages such as low sensitivity, complicated operation, and short storage time of reagents; enzyme immunoassay has low sensitivity and low signal-to-noise ratio. Methodological constraints such as narrow linear range; time-resolved fluorescence immunoassay has high requirements on the detection environment and is easily disturbed by air dust; chemiluminescence immunoassay is an advanced and effective method, which can make the detection sensitivity reach 10 -18 Molar level, and the detection range can reach 6 orders of magnitude, the enzyme label is stable and can be used for a long time

Method used

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  • C-peptide micropore plate type magnetic granule chemoluminescence immunoassay measuring kit and preparation method thereof
  • C-peptide micropore plate type magnetic granule chemoluminescence immunoassay measuring kit and preparation method thereof

Examples

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Effect test

Embodiment 1

[0059] Example 1 Preparation of C Peptide Microwell Plate Magnetic Particle Chemiluminescent Immunoassay Assay Kit Luminescent Substrate Solution

[0060] According to the formula of chemiluminescent substrate solution A:

[0061] Sodium dihydrogen phosphate (NaH 2 PO 4 2H 2 O) 2.19g

[0062] Disodium hydrogen phosphate (Na 2 HPO 4 12H 2 O) 12.9g

[0063] Luminol 4g

[0064] Ethylene glycol diamine tetraacetic acid (EGTA) 0.2g

[0065] Glycerol 25ml

[0066] Acetaminophen 0.05g

[0067] Double distilled water to volume 1000ml

[0068] After the preparation is completed, polyethylene plastic bottles are used, and each bottle is divided into 10ml.

[0069] According to the formula of chemiluminescence substrate solution B solution:

[0070] Sodium dihydrogen phosphate (NaH 2 PO 4 2H 2 O) 2.19g

[0071] Disodium hydrogen phosphate (Na 2 HPO 4 12H 2 O) 12.9g

[0072] 30% hydrogen peroxide (H 2 o 2 ) 200ml

[0073] Sodium chloride (NaCl) 9g

[0074] Double ...

Embodiment 2

[0076] The preparation of embodiment 2 C peptide calibrator

[0077] Matrix fluid formulation according to the calibrator of the present invention

[0078] Tris (Tris) 6.06g

[0079] Bovine Serum Albumin (BSA) 1.5g

[0080] 0.1mol / L hydrochloric acid (HCL) 3.45ml

[0081] Proclin300 1ml

[0082] Cabbage Red 0.1ml

[0083] Double distilled water 1000ml

[0084] Prepare calibrator matrix solution, use this matrix solution to dilute the pure C-peptide antigen (>90%) into calibrator, the concentrations are 0.2ng / ml, 0.6ng / ml, 2ng / ml, 6ng / ml, 15ng / ml , plus matrix solution 0ng / ml, a total of 6 bottles, 1ml in each bottle, freeze-dried for storage.

Embodiment 3

[0085] Example 3 Preparation of C-peptide Antibody Magnetic Particles

[0086] Prepare according to the diluent formula of the C-peptide antibody-coated magnetic particles in the preparation method of the kit of the present invention:

[0087] Sodium dihydrogen phosphate (NaH 2 PO 4 2H 2 O) 2.19g

[0088] Disodium hydrogen phosphate (Na 2 HPO 4 12H 2 O) 12.9g

[0089] Bovine Serum Albumin (BSA) 10g

[0090] Gelatin 15g

[0091] Ethylene glycol 50ml

[0092] Deionized water to volume 1000ml

[0093] After the preparation is completed, it should be mixed well, and after standing for 30 minutes, observe with the naked eye that there are no crystals or precipitates. It should be tested with an electronic pH meter or neutral pH test paper. The pH should be controlled between 7.0-7.5, and the optimal pH is 7.2. Then dispense into polyethylene plastic bottles of appropriate volume.

[0094] When the particle size is 1-2 μm, when the amino group is attached to the surface ...

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Abstract

The invention discloses a C-peptide micropore plate type magnetic granule chemoluminescence immunoassay measuring kit and a preparation method thereof. The kit comprises: (1) a C-peptide calibration sample, (2) magnetic granules coated by a C-peptide monoclonal antibody, (3) another monoclonal antibody or a polyclonal antibody marked by horse radish peroxidase (HRP), (4) a micropore plate, (5) washing solution and (6) chemoluminescence substrate solution on which the enzyme acts. The preparation method for preparing the kit further comprises the steps of preparing the calibration sample, preparing the magnetic granules coated by the C-peptide antibody, marking the C-peptide antibody with the horse radish peroxidase (HRP), preparing the washing solution and preparing the chemoluminescence substrate solution. The kit is simple, convenient, fast, sensitive, stable and the like.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine. Specifically, the invention discloses a C-peptide microporous plate type magnetic particle chemiluminescence immunoassay assay kit and a preparation method thereof. Background technique [0002] Human C-peptide is a straight chain composed of 31 amino acids with a molecular weight of 3020. C-peptide is a connecting peptide released during the formation of insulin from proinsulin. C-peptide and insulin are secreted from B cells into the blood at equal molecular weights. C-peptide has no biological activity (some scholars also believe that C-peptide has biological activity), nor does it bind to receptors on the cell membrane and is not degraded. Its half-life is 3-5 times that of insulin. Determination of C-peptide is an ideal indicator for diabetic patients treated with insulin. [0003] C-peptide is not inactivated by liver enzymes and is only excreted by the kidneys. It has a long half-...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N33/543G01N21/76
Inventor 王宏锐应希堂宋胜利胡国茂郑金来唐宝军张坤
Owner 北京科美东雅生物技术有限公司
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