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Preparation and application of a latex turbidimetric method for measuring c-inverse protein detection reagent

A technology for reacting proteins and latexes, which is applied in biological testing, analysis by chemical reaction of materials, and material analysis by observing the influence of chemical indicators, which can solve the problem of inaccurate determination of low-value samples and the amount of monoclonal antibody used. Large, high reagent costs, to achieve a wide detection range, improve detection sensitivity, good affinity

Active Publication Date: 2022-03-22
BIOSINO BIO TECH & SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the domestic double particle size latex microsphere method: the amount of monoclonal antibody used is large, the cost of reagents is high, and the sensitivity of most products in the low-value area is not high, and low-value samples (0.1-3.0mg / L) cannot be accurately determined

Method used

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  • Preparation and application of a latex turbidimetric method for measuring c-inverse protein detection reagent
  • Preparation and application of a latex turbidimetric method for measuring c-inverse protein detection reagent
  • Preparation and application of a latex turbidimetric method for measuring c-inverse protein detection reagent

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Experimental program
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Embodiment 1

[0059] The preparation of buffer solution in embodiment 1 reagent R2

[0060] Prepare various types of buffer

[0061] (1) Activation solution: 2-morpholineethanesulfonic acid (MES) concentration is 10mmol / L, pH: 6.00;

[0062] (2) Coupling buffer: sodium dihydrogen phosphate, the concentration is 10mmol / L, pH: 7.60;

[0063] (3) blocking solution: glycine buffer solution, concentration is 10mmol / L, bovine serum albumin (BSA) content: 200mg / L, pH:7.20;

[0064] (4) Washing buffer: glycine buffer, concentration is 10mmol / L, pH: 7.60;

[0065] (5) Preservation solution: the concentration of 4-hydroxyethylpiperazineethanesulfonic acid (HEPES) is 50mmol / L, pH: 8.10; in addition to HEPES, the preservation solution can also add: sucrose: 50g / L, mannitol: 50g / L, glycerin: 50g / L, trehalose: 20g / L, 0.1% Proclin-300.

Embodiment 2

[0067] 1. Preparation of CRP antibody-large particle size latex microsphere solution

[0068] (1) Activation of latex microspheres

[0069] Add latex microspheres in the activation solution, large particle size latex microspheres (220nm), the microsphere content is 0.35% of the activation solution volume, continue to add dissolved N-hydroxysuccinimide, carbodiimide in the activation solution Solution, so that the final concentration of NHS is 0.3mmol / L, and the final concentration of NHS is 1.0mmol / L, mix well at room temperature for 20 minutes, centrifuge and discard the supernatant solution to obtain activated latex microspheres;

[0070] (2) Coupling of latex microspheres and CRP antibody

[0071] Add the activated latex microspheres in (1) to the coupling buffer, resuspend the activated latex microspheres in the coupling buffer, ultrasonically mix, and add the activated latex microspheres to the coupling buffer containing the activated latex microspheres. Slowly add CRP ...

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Abstract

The invention relates to the preparation and application of a latex turbidimetric method for measuring C-inverse protein detection reagent and belongs to the field of in vitro diagnostic detection reagents. The detection reagent provided by the invention includes latex microspheres with large particle size and latex microspheres with small particle size; the C-reactive protein is detected by using the conjugate of latex microspheres with large particle size and CRP antibody. Using the CRP antibody-small particle size latex microspheres and CRP antibody-large particle size latex microspheres provided by the present invention can detect C-reactive protein in a full range, and the detection results have high sensitivity and specificity.

Description

technical field [0001] The invention belongs to the field of in vitro diagnostic detection reagents, in particular to the preparation and application of a latex turbidimetric C-reactive protein detection reagent. Background technique [0002] C-reactive protein (CRP) is an acute phase protein synthesized by liver cells when the body is subjected to inflammatory stimuli such as microbial invasion or tissue damage, and is a non-specific inflammatory marker. The content in the serum of healthy people is very low. When the human body is attacked by various factors such as microorganisms, an acute phase reaction protein will be produced in the human serum a few hours later, and this protein hyperreacts with the C polysaccharide of pneumococcus. Structurally, CRP contains 5 polypeptide chain subunits, which are non-covalently combined into a disc-shaped polymer with a molecular weight of 115,000-140,000. It is a typical acute phase protein. [0003] CRP detection kits can be divi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/543G01N21/82
CPCG01N33/6893G01N33/54346G01N21/82G01N2333/4737
Inventor 李珂夏另朝
Owner BIOSINO BIO TECH & SCI