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preparation method and application of C-reactive protein immuno latex microsphere

A technology of reacting proteins and microspheres, applied in the field of medical immunodiagnosis, can solve the problems of low product yield and little effect on antibody activity of coupling efficiency, and achieve the effects of high sensitivity, good clinical application prospects and fast operation.

Active Publication Date: 2017-01-11
WUHAN KING DIAGNOSTIC TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to overcome the defect of low product yield in the existing C-reactive protein immune latex microsphere preparation method, and provide a C-reactive protein immune latex microsphere with high coupling efficiency and little influence on antibody activity after coupling Preparation method and C-reactive protein assay kit by latex-enhanced immune turbidimetric method using the C-reactive protein immune latex microspheres

Method used

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Embodiment 1

[0051] The preparation method of C-reactive protein immune latex microspheres provided by the present invention is as follows:

[0052] 1. Prepare polystyrene microspheres with a particle size of 120nm, the concentration is 10%, and take 5ml; the polystyrene microspheres are first diluted with 2-morpholineethanesulfonic acid buffer solution with a concentration of 50mmol / L to a final concentration of 1%. w / v, to obtain a microsphere solution with a pH of 5.5 to 6.5, the volume of the solution is 50 mL;

[0053] 2. Dissolve carbodiimide in 2-morpholineethanesulfonic acid buffer to obtain a carbodiimide solution with a concentration of 10 mg / mL;

[0054] 3. Based on the weight of polystyrene microspheres contained in the microsphere solution, add the carbodiimide solution dropwise to the microsphere solution at a ratio of 50-60 μL per 10 mg of polystyrene microspheres, and add dropwise 250 μL, shake and activate at 4°C for 1 hour at a speed of 50 r / min to obtain an activated po...

Embodiment 2

[0058] The preparation method of C-reactive protein immune latex microspheres provided by the present invention is as follows:

[0059] 1. Prepare polystyrene microspheres with a particle size of 50nm, the concentration is 10%, take 5ml; the polystyrene microspheres are first diluted with 2-morpholineethanesulfonic acid buffer solution with a concentration of 50mmol / L to a final concentration of 0.8% w / v, to obtain a microsphere solution with a pH of 5.5 to 6.5, the volume of the solution is 50 mL;

[0060] 2. Dissolve carbodiimide in 2-morpholineethanesulfonic acid buffer to obtain a carbodiimide solution with a concentration of 10 mg / mL;

[0061] 3. Based on the weight of polystyrene microspheres contained in the microsphere solution, add the carbodiimide solution dropwise to the microsphere solution at a ratio of 50-60 μL per 10 mg of polystyrene microspheres, and add dropwise 240 μL, shake and activate at 8°C for 1 hour at a speed of 30 r / min to obtain an activated polyst...

Embodiment 3

[0065] The preparation method of C-reactive protein immune latex microspheres provided by the present invention is as follows:

[0066] 1. Prepare polystyrene microspheres with a particle size of 200nm, the concentration is 10%, take 5ml; the polystyrene microspheres are first diluted with 2-morpholineethanesulfonic acid buffer solution with a concentration of 50mmol / L to a final concentration of 1.2% w / v, to obtain a microsphere solution with a pH of 5.5 to 6.5, the volume of the solution is 50 mL;

[0067] 2. Dissolve carbodiimide in 2-morpholineethanesulfonic acid buffer to obtain a carbodiimide solution with a concentration of 10 mg / mL;

[0068] 3. Based on the weight of polystyrene microspheres contained in the microsphere solution, add the carbodiimide solution dropwise to the microsphere solution at a ratio of 50-60 μL per 10 mg of polystyrene microspheres, and add dropwise 300 μL, shake and activate at 2°C for 1 hour at a speed of 50 r / min to obtain an activated polys...

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Abstract

The invention discloses a preparation method and application of C-reactive protein immuno latex microsphere. The preparation method of C-reactive protein immuno latex microsphere optimizes a chemical crosslinking method. The carboxyl of polystyrene microspheres is activated at a low pH, and is coupled with the amino group of a dissolved antibody at a high pH, the mixture of polystyrene microspheres and the dissolved antibody is neutral, after coupling for 12-24h at low temperature, the coupling efficiency of the antibody is ensured, and the damage of an activating agent on antibody activity is greatly reduced. A latex enhanced immunoturbidimetry C-reactive protein determination kit prepared by the immuno latex microsphere which is produced by the C-reactive protein immuno latex microsphere preparation method has the advantagess of high sensitivity, high accuracy, good repeatability and stable property, the quantitative linear range is 0.1-20 mg / L, and the latex enhanced immunoturbidimetry C-reactive protein determination kit can be used in a full-automatic biochemical analyzer or a coagulation analyzer. The operation is fast and simple, it only takes 5 to 10 minutes from detection to result collection, and the prospect for clinical application is good.

Description

technical field [0001] The invention relates to the field of medical immunodiagnosis, in particular to a preparation method and application of C-reactive protein immune latex microspheres. Background technique [0002] C-reactive protein (C-reactive protein, CRP) is an acute-phase reactive protein mainly produced by the liver under the mediation of interleukin-6, and its concentration in the acute phase can increase by thousands of times. [0003] CRP is one of the important proteins in the acute phase response proteins in humans and animals, and it is a sensitive inflammatory marker, and its blood concentration rises sharply during acute trauma and infection. Normal animal serum CRP content is very low, but in the case of infection, inflammation, surgery and tumor, etc., it rises rapidly in a few hours, and reaches the peak in 24 to 48 hours in dogs. After the lesions subside, CRP can quickly drop to normal, so it can be used as a preferred indicator of bacterial and viral...

Claims

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Application Information

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IPC IPC(8): G01N33/531G01N33/68
CPCG01N33/531G01N33/6893G01N2333/4737
Inventor 王明吴艳芳陈莹
Owner WUHAN KING DIAGNOSTIC TECH CO LTD
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