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Trypsin micropore plate type magnetic granule chemoluminescence immunoassay measuring kit and preparation method thereof

A chemiluminescence immunoassay and insulin technology, applied in the field of immunoassay medicine, can solve problems such as being easily interfered by air dust, radioactive pollution in the environment, narrow linear range, etc., and achieve strong clinical applicability, no radioactive pollution, and wide detection range 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

[0004] 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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  • Trypsin micropore plate type magnetic granule chemoluminescence immunoassay measuring kit and preparation method thereof
  • Trypsin micropore plate type magnetic granule chemoluminescence immunoassay measuring kit and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0054] Example 1 Preparation of insulin antibody magnetic particles

[0055] Prepare according to the diluent formula of the insulin antibody-coated magnetic particles in the preparation method of the kit of the present invention:

[0056] Tris (Tris) 6.06g

[0057] 0.1mol / L hydrochloric acid (HCL) 3.45ml

[0058] Bovine Serum Albumin (BSA) 10g

[0059] Gelatin 15g

[0060] Ethylene glycol 50ml

[0061] Deionized water to volume 1000ml

[0062] 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 7.0-8.0, and the optimal pH is 7.8. Then dispense into polyethylene plastic bottles of appropriate volume.

[0063] When the particle size is 1-2 μm, when the amino group is attached to the surface of the magnetic particle, the magnetic particle is treated wi...

Embodiment 2

[0064] Example 2 Preparation of Washing Solution in Insulin Kit

[0065] Prepare according to the formula of washing liquid in the kit preparation method of the present invention:

[0066] Tris (Tris) 1.212g

[0067] 0.1mol / L hydrochloric acid (HCL) 0.69ml

[0068] Tween20 0.1ml

[0069] Deionized water to volume 1000ml

[0070] 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 7.0-8.0, and the optimal pH is 7.8. Then dispense the washing solution into polyethylene plastic bottles of appropriate size.

Embodiment 3

[0071] Example 3 Preparation of Luminescent Substrate Solution of Insulin Microwell Plate Magnetic Particle Chemiluminescence Immunoassay Assay Kit

[0072] According to the formula of chemiluminescent substrate solution:

[0073] Tris (Tris) 6.06g

[0074] 0.1mol / L hydrochloric acid (HCL) 3.45ml

[0075] 4-Methoxy-4-(3-phosphoryloxyphenyl)-helix-(1,2-dioxetane-3,2′-adamantane)

[0076] 100ml

[0077] Magnesium chloride 0.02g

[0078] Double distilled water to volume 1000ml

[0079] After the preparation is completed, polyethylene plastic bottles are used, and each bottle is divided into 10ml.

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Abstract

The invention discloses a trypsin micropore plate type magnetic granule chemoluminescence immunoassay measuring kit and a preparation method thereof. The kit comprises: (1) a trypsin calibration sample, (2) magnetic granules coated by a trypsin monoclonal antibody, (3) another monoclonal antibody or a polyclonal antibody marked by alkaline phosphatase (ALP), (4) a micropore plate, (5) washing solution and (6) chemoluminescence substrate solution on which the enzyme acts. The preparation method for preparing the kit comprises the steps of preparing the calibration sample, preparing the magnetic granules coated by the trypsin antibody, marking the trypsin antibody with the alkaline phosphatase (ALP), preparing the washing solution and preparing the chemoluminescence substrate solution. The kit can carry out batch measurement and is suitable for batch clinic blood serum screening.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine. Specifically, the invention discloses an insulin microporous plate type magnetic particle chemiluminescence immunoassay assay kit and a preparation method thereof. Background technique [0002] Insulin has a molecular weight of 5700 and consists of two amino acid peptide chains. The A chain has 21 amino acids and the B chain has 30 amino acids. Among them, the sulfhydryl groups in the four cysteines of A7(Cys)-B7(Cys) and A20(Cys)-B19(Cys) form two disulfide bonds to connect the two chains of A and B. In addition, there is also a disulfide bond between A6(Cys) and A11(Cys) in the A chain. On an empty stomach, the plasma insulin concentration is 5-15 μIU / mL. Plasma insulin levels can increase 5-10 times after a meal. The biosynthesis rate of insulin is affected by the plasma glucose concentration. When the blood glucose concentration rises, the content of proinsulin in B cells increases,...

Claims

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Application Information

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IPC IPC(8): G01N33/74G01N33/577G01N33/543G01N21/76
Inventor 王宏锐应希堂宋胜利胡国茂郑金来唐宝军于尚永
Owner 北京科美东雅生物技术有限公司
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