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A kind of preparation method of chloromethylated polystyrene latex

A polystyrene latex and chloromethylation technology, applied in the field of immunoassay, can solve the problems of few research reports, health hazards of operators, easy variation, etc., and achieve the effects of simple process method, long storage time and simple operation.

Active Publication Date: 2020-06-19
WUHAN LIFE ORIGIN BIOTECH LTD
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  • Description
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AI Technical Summary

Problems solved by technology

[0003] At present, the latex particles used in LETIA detection reagents are mostly carboxylated microspheres. The combination of surface-modified latex particles and antibodies is through the covalent bonding of the carboxyl groups on the surface with the amino terminals of antibodies, but the storage time of carboxylated microspheres is relatively short. , easy to vary; there are many reports on carboxylated microspheres, but there are fewer reports on chloromethylated microspheres. In the existing technology for preparing chloromethylated, chloromethyl ether is often used to chlorinate polystyrene resins. Methylation, that is, under the action of a catalyst, it is produced by the direct or indirect reaction of polystyrene cross-linked microspheres and chloromethyl ether, but chloromethyl ether has serious carcinogenic toxicity and is easy to cause health hazards to operators, so it is not suitable for long-term exposure area use

Method used

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  • A kind of preparation method of chloromethylated polystyrene latex
  • A kind of preparation method of chloromethylated polystyrene latex

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] 1.1 Preparation of chloromethylated polystyrene latex

[0019] (1) Draw 100 mL of polystyrene latex particle suspension, wash and dry, and weigh to obtain 1 g of latex particle;

[0020] (2) Take paraformaldehyde twice the weight of polystyrene latex and 30% hydrochloric acid twice the weight, mix and stir, heat up to 65°C and reflux for 3 hours, the temperature gradually rises to about 85°C, after the reflux is completed, cool to 4°C.

[0021] (3) add weighed polystyrene latex particles in the reaction solution after reflux, after stirring evenly, add manganese dichloride of 0.1 times of weight of latex particles as a catalyst, after being incubated at 25°C for 10 minutes, add acetic anhydride and Chlorocyclohexane was post-mixed and reacted for 3 hours.

[0022] (4) Centrifuge, discard the solution, add 10% hydrochloric acid and paraformaldehyde mixed solution to the latex, and store in the dark for future use.

[0023] 1.2 Immediately after the preparation of chlor...

Embodiment 2

[0033] 2.1 Preparation of chloromethylated polystyrene latex

[0034] (1) absorb polystyrene latex particle suspension, weigh after washing with water;

[0035] (2) Mix and stir paraformaldehyde with 1 times the weight of polystyrene latex and 10% hydrochloric acid with 1 times the weight of polystyrene latex, heat up to 65°C and reflux for 5 hours, the temperature gradually rises to about 90°C, after the reflux ends, cool to 4°C;

[0036] (3) add weighed polystyrene latex particles in the reaction solution after reflux, after stirring evenly, add manganese dichloride of 0.1 times of weight of latex particles as a catalyst, after being incubated at 25°C for 10 minutes, add acetic anhydride and Chlorocyclohexane was mixed and reacted for 3 hours;

[0037] (4) Centrifuge, discard the solution, add 10% hydrochloric acid and paraformaldehyde mixed solution to the latex, and store in the dark for future use.

[0038] 2.2 Immediately after the preparation of chloromethylated poly...

Embodiment 3

[0047] Embodiment 3: comparative example

[0048] 3.1 Carboxyl latex-conjugated antibody just prepared by prior art

[0049] The operation steps are as follows, taking 4mL activated latex-conjugated antibody as an example:

[0050] (1) Preparation of reagents

[0051] ①. Prepare 50mM MES buffer (activation) with pH=5.0;

[0052] ②. Prepare 50mM MES buffer (coupling) with pH=6.0;

[0053] ③. Prepare 100mg / ml NHS solution;

[0054] ④. Prepare 50mg / ml EDAC solution;

[0055] ⑤ Latex: NHS:EDAC=10:15:3;

[0056] ⑥ Cleaning solution: 1mM HCl, 100ml, pre-cooled at 4°C;

[0057] ⑦. Coupling buffer: 0.2M NaHCO3+0.5M NaCl, pH 8.3, 50ml;

[0058] (2) Activated carboxyl latex:

[0059] ①. Add 690 μL of pH5.0 50mM MES buffer into a 1.5mL centrifuge tube;

[0060] ②. Add 100 μL of 10% latex, and vortex to mix;

[0061] ③. Add 150 μL of NHS solution;

[0062] ④. Add 60 μL of EDAC and vortex to mix;

[0063] ⑤, constant temperature (30 ℃) shaker activation 200rpm 30min;

[0064] ⑥....

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Abstract

The invention provides a preparation method of chloromethylated polystyrene latex. The method includes the steps of: sucking a polystyrene latex particle suspension, conducting washing and drying to obtain latex particles, and performing weighing; mixinging paraformaldehyde with 30% hydrochloric acid, wherein the paraformaldehyde, polystyrene latex particles and hydrochloric acid are in a weight ratio of 1-2:1:1-2; adding polystyrene latex particles, stirring the substances evenly, then adding manganese chloride that is 0.1 times the weight of the polystyrene latex particles, performing heat preservation at 25DEG C for 8-15min, then adding a small amount of acetic anhydride and chlorocyclohexane, carrying out mixing reaction for 2-4h, performing centrifugation and rejecting the solution, adding 10% hydrochloric acid and a trioxymethylene mixed solution, and conducting storage in dark place for standby use. The technological method provided by the invention is simple, the reactants andreaction product have small toxicity, and the prepared chloromethylated polystyrene latex has better stability and longer preservation time compared with carboxylated microspheres, and can directly used for antibody coupling without activation.

Description

technical field [0001] The invention relates to the field of immune analysis, in particular to a preparation method of chloromethylated polystyrene latex. Background technique [0002] Generally, in the field of immunoassays, the antigen-antibody reaction is first performed to form an immune complex, and then quantified by turbidimetric method, but it takes a long time to form the antigen-antibody complex. Latex Enhanced Immunoturbidimetric Assay (LETIA for short) is a new immunoassay technology that has emerged in the field of immunoassays in recent years. By coupling latex to antibodies, the reactivity of the antibodies can be changed and the immunoprecipitation reaction can be enhanced. Latex-enhanced immune turbidimetry, the principle is that latex particles (immune latex) coated with antigens or antibodies, after an immune reaction with the corresponding antibodies or antigens in the specimen, form agglutinated particles, and the formation of agglutinates makes the reac...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08F8/24C08F12/08G01N33/543
CPCC08F8/24G01N33/54313C08F12/08
Inventor 华权高来祥兵徐春雷
Owner WUHAN LIFE ORIGIN BIOTECH LTD
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