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Method and reagent for latex sensitization

A latex and reagent technology, applied in the field of latex-enhanced immune turbidimetric reagents, can solve problems such as long time, loss of antigen or antibody activity, sensitivity impact, etc., to save time, improve detection sensitivity, and enhance clinical application value.

Active Publication Date: 2011-08-24
BEIJING MDC NEW SPRING MEDICAL DEVICES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the chemical cross-linking method will cause the loss of activity of the antigen or antibody during the cross-linking process due to chemical reactions, which will affect the sensitivity of the reagents.
The usual steps of the chemical cross-linking method are to directly cross-link the antigen or antibody on the latex through chemical bonds to make it sensitized, and then add BSA, polyethylene glycol and other proteins or polymers to block the unbound sites on the latex surface , the whole reaction generally takes two days or more, and the time is longer
Moreover, because the reaction conditions are strong and the reaction time is long, the activity of the antigen or antibody is greatly lost, and the sensitivity of the reaction is also greatly affected.

Method used

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  • Method and reagent for latex sensitization
  • Method and reagent for latex sensitization
  • Method and reagent for latex sensitization

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Binding of Latex to Unrelated Proteins

[0046] raw material:

[0047] Carboxyl-modified polystyrene latex, particle diameter 124nm, concentration 10% (weight to volume ratio), surface carboxyl content 100 μ mol / g.

[0048] EDAC (analytical grade, sigma) was dissolved in double-distilled water to prepare a 5 mg / ml solution, which was prepared half an hour before use.

[0049] With 10mM MES, pH6.1 damping fluid is diluted latex solution to 0.5% (weight volume ratio), then adds BSA solution, makes the concentration of BSA in the system reach 200ug / ml, finally adds the EDAC solution of 5mg / ml, makes EDAC When the concentration in the system reaches 75ug / ml, the reaction starts. A constant temperature water bath was used to keep the reaction temperature at 37° C. and the reaction time was 3 hr. After the reaction, centrifuge (12000rpm, 30min), then add the initial same amount of 0.1M PBS, pH7.4 buffer solution, and ultrasonically in ice bath to disperse the la...

Embodiment 2

[0050] Embodiment 2: Preparation of sensitized latex

[0051] To the unrelated protein-bound latex solution in Example 1 was added streptolysin O antigen. The streptolysin O antigen used is a gene recombination product. The concentration of the added antigen in the system was 100ug / ml. Then add 2% glutaraldehyde aqueous solution to start the reaction, the addition ratio is 0.002%. The reaction temperature is 25°C, the reaction time is 3hr, after the reaction is over, centrifuge (12000rpm, 30min), then add the same amount of 0.1M glycine at the beginning, pH8.4 buffer solution, ice bath ultrasonication, to disperse the latex, centrifuge again, and the latex Disperse in 0.1M glycine containing 0.5% (weight to volume ratio) BSA, and store in pH 8.4 buffer.

Embodiment 3

[0052] Example 3: Preparation of Antistreptolysin O Latex Enhanced Immunoturbidimetric Reagent

[0053] The specific composition of antistreptolysin O latex enhanced immune turbidimetric reagent is as follows:

[0054] Reagent R1: Glycine buffer 0.1M pH8.4

[0055] NaCl 300mM

[0056] Tween 20 0.1% (weight to volume ratio)

[0057] PEG6000 1% (weight to volume ratio)

[0058] NaN 3 0.1% (weight to volume ratio)

[0059] Reagent R2: Glycine buffer 0.1M pH8.4

[0060] Allergen latex (prepared in embodiment 2) 0.17% (weight to volume ratio)

[0061] BSA 0.5% (weight to volume ratio)

[0062] NaN 3 0.1% (weight to volume ratio).

[0063] The control is a commercially available imported reagent A, the information of which is as follows:

[0064] Product Name: Anti-Streptococcal O Detection Kit (Immunoturbidimetric Assay)

[0065] Principle: The ASO in the sample reacts with the hypersensitized latex particle reagent to form an immune complex, and the c...

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Abstract

The invention relates to a method and a reagent for latex sensitization. Specifically, the method for latex sensitization comprises the following steps: combining latex with an unrelated protein by a chemical bond; and then, crosslinking an antigen or an antibody on the unrelated protein by glutaraldehyde so as to cause latex sensitization. The antistreptolysin O latex prepared with the method enhances an immune turbidimetry reagent. With the method disclosed by the invention, time for preparing the reagent can be shortened, and the sensitivity of the reagent is improved.

Description

technical field [0001] The invention relates to a method and a reagent for latex sensitization. More particularly, the present invention relates to a method for sensitizing latex by first combining latex with unrelated proteins through chemical bonds, and then using glutaraldehyde to cross-link antigens or antibodies to unrelated proteins. The invention also relates to the sensitized latex obtained by the method and the latex-enhanced immune turbidimetric reagent using the sensitized latex. Background technique [0002] In 1956, Singer and Plotz began to use particles with a diameter of microns to perform qualitative slide agglutination detection of antibodies or antigens in serum through antigens or antibodies attached to latex. The latex-enhanced immunoturbidimetric method developed on this basis improves the sensitivity of the ordinary immunoturbidimetric method, and is easy to operate and can be determined by ordinary biochemical analyzers. It is a new clinical diagnost...

Claims

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Application Information

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IPC IPC(8): C08F12/08C08F8/30C08G81/02C08H1/00G01N33/53
Inventor 胥敏刘希高爱民
Owner BEIJING MDC NEW SPRING MEDICAL DEVICES
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