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Glycosylated hemoglobin detection reagent and latex microsphere and polylysine coupling method

A technology of glycosylated hemoglobin and polylysine, which is applied in the field of biochemical analysis to achieve the effects of improving detection sensitivity, improving efficiency, and shortening reaction and detection time

Pending Publication Date: 2020-06-05
SUZHOU DIAGVITA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The object of the present invention is to provide a kind of glycosylated hemoglobin detection reagent which can solve the aforementioned problems in view of the problems of the existing glycosylated hemoglobin detection reagents described in the background technology

Method used

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  • Glycosylated hemoglobin detection reagent and latex microsphere and polylysine coupling method
  • Glycosylated hemoglobin detection reagent and latex microsphere and polylysine coupling method
  • Glycosylated hemoglobin detection reagent and latex microsphere and polylysine coupling method

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Experimental program
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Effect test

Embodiment 1

[0020] Glycated hemoglobin detection reagent, it comprises R1 reagent, R2 reagent and R3 reagent, in R1 reagent, the particle diameter of polystyrene latex microsphere is 50nm, and concentration is 0.05% (mass / volume ratio), the concentration of polylysine 0.1g / L, the molecular weight is 1000, the coupling steps of latex microspheres and poly-lysine are as follows:

[0021] 1) Measure 0.5mL of 50nm latex microspheres (10%), place in a 10ml beaker, and use a medium-sized stirrer bar for magnetic stirring;

[0022] 2) Slowly add 0.1ml 500mM MES BUFFER (pH6.0);

[0023] 3) Weigh 50mg EDAC, 5mg Sulfo-NHS, and 0.01g polylysine respectively, and add them to the beaker;

[0024] 4) room temperature, magnetic stirring for 1 hour;

[0025] 5) Transfer the latex microspheres to a 50ml centrifuge tube, rinse the beaker with 10ml deionized water, and transfer to the centrifuge tube together;

[0026] 6) Centrifuge after trimming, 16000rpm, 8 degrees, 60 minutes;

[0027] 7) Pour off t...

Embodiment 2

[0034] Glycated hemoglobin detection reagent, it comprises R1 reagent, R2 reagent and R3 reagent, in R1 reagent, the particle diameter of polystyrene latex microsphere is 100nm, and concentration is 0.2% (mass / volume ratio), the concentration of polylysine Be 1g / L, molecular weight is 5000, latex microsphere and polylysine coupling:

[0035] a. Measure 2mL of 100nm latex microspheres (10%), place in a 10ml beaker, and use a medium-sized stirring bar to stir magnetically;

[0036] b. Slowly add 0.1ml 500mM MES BUFFER (pH6.0);

[0037] c. Weigh 50mg EDAC, 5mg Sulfo-NHS, and 0.1g polylysine respectively, and add them to the beaker;

[0038] d. Stir magnetically for 1 hour at room temperature;

[0039] e. Transfer the latex microspheres to a 50ml centrifuge tube, rinse the beaker with 10ml deionized water, and transfer to the centrifuge tube together;

[0040] f. Centrifuge after trimming, 16000rpm, 8 degrees, 60 minutes;

[0041] g. Pour off the supernatant, add 5ml deionized...

Embodiment 3

[0048] Glycated hemoglobin detection reagent, it comprises R1 reagent, R2 reagent and R3 reagent, in R1 reagent, the particle diameter of polystyrene latex microsphere is 300nm, and concentration is 0.3% (mass / volume ratio), the concentration of polylysine Be 2g / L, molecular weight is 10000, latex microsphere is coupled with polylysine:

[0049] a. Measure 3mL 300nm latex microspheres (10%), place in a 10ml beaker, and use a medium-sized stirring bar to stir magnetically;

[0050] b. Slowly add 0.1ml 500mM MES BUFFER (pH6.0);

[0051] c. Weigh 50mg EDAC, 5mg Sulfo-NHS, and 0.2g polylysine respectively, and add them to the beaker;

[0052] d. Stir magnetically for 1 hour at room temperature;

[0053]e. Transfer the latex microspheres to a 50ml centrifuge tube, rinse the beaker with 10ml deionized water, and transfer to the centrifuge tube together;

[0054] f. Centrifuge after trimming, 16000rpm, 8 degrees, 60 minutes;

[0055] g. Pour off the supernatant, add 5ml deionized...

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Abstract

The invention discloses a glycosylated hemoglobin detection reagent. The reagent comprises a reagent R1, a reagent R2 and a reagent R3, the R2 reagent comprises a primary antibody and a secondary antibody; the primary antibody is a mouse anti-human HbA1c monoclonal antibody; the first antibody is a glycosylated hemoglobin monoclonal antibody, the second antibody is a goat anti-mouse IgG polyclonalantibody, the R3 reagent comprises a hemolytic agent, R1 is a polystyrene latex reagent, latex microspheres of the R1 reagent are coupled with polylysine, the binding adsorption positions of the surfaces of the latex microspheres and glycosylated hemoglobin are increased through coupling of the polylysine and the latex microspheres, and the binding efficiency is improved. According to the invention, polylysine is coupled to latex microspheres of the reagent R1; through coupling of polylysine and latex microspheres, the binding adsorption positions of the surfaces of the latex microspheres andglycosylated hemoglobin are increased, the binding efficiency of the surfaces of the latex microspheres and the glycosylated hemoglobin can be improved, the detection sensitivity of the glycosylatedhemoglobin detection reagent is obviously improved, the reaction detection time is shortened, and the detection efficiency is improved.

Description

technical field [0001] The invention relates to the technical field of biochemical analysis, in particular to a detection reagent for glycosylated hemoglobin and a coupling method between latex microspheres and polylysine. Background technique [0002] At present, the detection methods of glycosylated hemoglobin that are widely used clinically are mainly high-performance liquid chromatography, affinity chromatography, electrophoresis, etc. These detection methods are complicated to operate, time-consuming, and require special instruments, and the cost is high. There are also other detection methods in the world, but they generally have the defect of low measurement accuracy. [0003] The latex particle-enhanced turbidimetric method is a relatively stable and accurate method for homogeneous immune turbidimetric detection of body fluid proteins that has emerged in recent years. The specific principle is to cross-link or physically adsorb antibodies on the surface of polymer l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/72G01N33/577G01N33/543
CPCG01N33/54346G01N33/577G01N33/723
Inventor 赵年福张辉贾引军王明伟金君玉吴一凡
Owner SUZHOU DIAGVITA BIOTECH
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