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Preparation method and application of d-dimer immune latex microspheres

A dimer, polystyrene microsphere technology, applied in the field of medical immunodiagnosis, can solve the problems of low product yield, little effect of coupling efficiency and antibody activity, and achieve high sensitivity, good clinical application prospects, and reduced damage. Effect

Active Publication Date: 2018-06-19
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 D-dimer immune latex microsphere preparation method, to provide a D-dimer immune latex with high coupling efficiency and little influence on antibody activity after coupling Microsphere preparation method and D-dimer determination kit by latex-enhanced immune turbidimetric method using the D-dimer immune latex microspheres

Method used

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  • Preparation method and application of d-dimer immune latex microspheres

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Embodiment 1

[0054] The preparation method of D-dimer immune latex microspheres provided by the present invention is as follows:

[0055] 1. Prepare polystyrene microspheres with a particle size of 240nm, the concentration is 10%, take 2ml; the polystyrene microspheres are first diluted with 2-morpholineethanesulfonic acid buffer solution with a concentration of 50mmol / L to a final concentration of 0.5% w / v, to obtain a microsphere solution with a pH of 5.5 to 6.5, the volume of the solution is 40mL;

[0056] 2. Dissolve carbodiimide in 2-morpholineethanesulfonic acid buffer to obtain a carbodiimide solution with a concentration of 10 mg / mL;

[0057] 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 35-45 μL per 10 mg of polystyrene microspheres, and add dropwise 80 μL, shake and activate at 4°C for 1 hour at a shaking speed of 50 r / min to obtain an activated polystyren...

Embodiment 2

[0061] The preparation method of D-dimer immune latex microspheres provided by the present invention is as follows:

[0062] 1. Prepare polystyrene microspheres with a particle size of 200nm, the concentration is 10%, take 2ml; the polystyrene microspheres are first diluted with 2-morpholineethanesulfonic acid buffer solution with a concentration of 50mmol / L to a final concentration of 0.4% w / v, to obtain a microsphere solution with a pH of 5.5 to 6.5, the volume of the solution is 50 mL;

[0063] 2. Dissolve carbodiimide in 2-morpholineethanesulfonic acid buffer to obtain a carbodiimide solution with a concentration of 10 mg / mL;

[0064] 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 35-45 μL per 10 mg of polystyrene microspheres, and add dropwise 80 μL, shake and activate at 8°C for 1 hour at a speed of 30 r / min to obtain an activated polystyrene micro...

Embodiment 3

[0068] The preparation method of D-dimer immune latex microspheres provided by the present invention is as follows:

[0069] 1. Prepare polystyrene microspheres with a particle size of 400nm, the concentration is 10%, take 2ml; the polystyrene microspheres are first diluted with 2-morpholineethanesulfonic acid buffer solution with a concentration of 50mmol / L to a final concentration of 0.6% w / v, to obtain a microsphere solution with a pH of 5.5 to 6.5, the volume of the solution is 40mL;

[0070] 2. Dissolve carbodiimide in 2-morpholineethanesulfonic acid buffer to obtain a carbodiimide solution with a concentration of 10 mg / mL;

[0071] 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 35-45 μL per 10 mg of polystyrene microspheres, and add dropwise 84 μL, shake and activate at 2°C for 1 hour at a speed of 50 r / min to obtain an activated polystyrene micros...

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Abstract

The invention discloses a preparation method and application of D-dimer immuno latex microspheres. In the preparation method of D-dimer immuno latex microspheres, a chemical crosslinking method is optimized, the carboxyl of polystyrene microsphere is activated in a low-pH condition and then coupled with amino of an antibody dissolved in a high-pH condition, and the mixture of the two is neutral and is coupled for 12-24h at a low temperature, so that not only can the coupling efficiency of the antibody be guaranteed, but also the damage of the antibody activity caused by an activator is reduced to a great extent. A latex enhanced turbidimetric immunoassay D-dimer test kit prepared from the immuno latex microspheres prepared by the method has the characteristics of high sensitivity, accurate quantification, good repeatability and stable property, and the lowest detection limit can reach 0.01mg / L; the latex enhanced turbidimetric immunoassay D-dimer test kit can be applied to a full-automatic biochemical analyzer or coagulation analyzer, is quick and easy to operate, only needs 5-10 minutes from detection to result collection and has relatively good clinical application prospect.

Description

technical field [0001] The invention relates to the field of medical immunodiagnosis, in particular to a preparation method and application of D-dimer immune latex microspheres. Background technique [0002] D-dimer is a specific degradation product produced by fibrin monomer cross-linked by activated factor XIII and then hydrolyzed by plasmin, and is a specific marker of fibrinolysis process. [0003] Plasma D-dimer measurement is a test to understand the function of secondary fibrinolysis. The principle of the latex-enhanced immune turbidimetric method used in the prior art is as follows: the anti-D-D monoclonal antibody is coated on the latex particles, if there is D-dimer in the recipient plasma, an antigen-antibody reaction will occur, and the latex particles will aggregate Phenomenon. [0004] In 1977, Sternberg created the rate-scattering turbidimetry method, which is to judge the content of antigen in the sample by the degree of scattering of light by the measured ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543
CPCG01N33/54346
Inventor 王明李华中吴艳芳
Owner WUHAN KING DIAGNOSTIC TECH CO LTD
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