CN101775094B - Preparation method for polystyrene latex with carboxyl on surface and for immunoassay technology - Google Patents

Preparation method for polystyrene latex with carboxyl on surface and for immunoassay technology Download PDF

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CN101775094B
CN101775094B CN2010101034280A CN201010103428A CN101775094B CN 101775094 B CN101775094 B CN 101775094B CN 2010101034280 A CN2010101034280 A CN 2010101034280A CN 201010103428 A CN201010103428 A CN 201010103428A CN 101775094 B CN101775094 B CN 101775094B
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邹炳德
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Medical System Biotechnology Co ltd
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Ningbo Medical System Biotechnology Co Ltd
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Abstract

The invention discloses a preparation method for polystyrene latex with carboxyl on the surface and for immunoassay technology; the preparation method comprises the following steps that: styrene, methyl methacrylate and water are added into a three-neck flask to be stirred for 10 to 30min; the molar ratio of the methyl methacrylate to the styrene is 0.1 to 0.2:1; after stirring, N2 is fed into the reaction system; and then potassium persulfate and the aqueous solution of sodium chloride are added in, so that the concentration of sodium chloride in the reaction system is 0.064mol/L, and the concentration of the potassium persulfate is 7.00*10-4 to 25.0*10-3mol/L; N2 is continued to be fed in; the temperature goes up to 60 to 80DEG C, and the reaction is carried out for 12 to 24h; and after the reaction, the temperature is cooled to room temperature, and the target product of the invention is prepared. The latex prepared by the invention has the advantages of uniform grain size, good monodispersity and strong stability; and simultaneously, the surface of the latex has higher carboxyl content, and the preparation method is applicable to a latex immune turbidity determination method.

Description

The preparation method of polystyrene latex who is used for the surface band carboxyl of immunoassay technology
Technical field
The present invention relates to a kind of preparation method of polystyrene latex of the surface band carboxyl that is used for immunoassay technology.
Background technology
In field of immunodetection generally speaking, at first antigen antibody reaction is formed immunocomplex, use the nephelometry quantitative assay then, need the long period but form antigen-antibody complex.Latex enhancing immune tuurbidimetry (being called for short LETIA) is a kind of new immunoassay technology of field of immunodetection appearance in recent years, LETIA does not need specific apparatus, automatically, semi-automatic biochemical analyzer can measure, suit to apply in vast basic medical treatment unit.Being applied to many analyte quantitative assays at present has tens of kinds more than, as ASO, RF, CRP, Transferrins,iron complexes, toxoplasma, rubella, β 2-microglobulin and drug surveillance etc.
Its principle of LETIA has been that bag is by the present latex particulate (immune latex) of antigen or antibody, after corresponding antibodies in the sample or antigen generation immune response, form agglutinating particle, the formation of agglutinator makes the reaction mixing system form certain turbidity, checking matter concentration is proportional in degree that turbidity increases and the sample, under certain wavelength, carry out turbidity measurement, can measure the content of checking matter in the sample.Therefore, how preparing suitable latex (immune latex) is the key of latex enhancing immune turbidity measurement.
At present, how conventional latex preparation process will use emulsifying agent as auxiliary material, but after latex preparation is finished, wherein tend to residual a certain amount of emulsifying agent, and the existence of emulsifying agent can influence the final use properties of latex.
In addition, the wavelength that the latex enhancing immune turbidity measurement is used is relevant with used latex particle size size, and the uniform particle diameter of the latex particle of prior art for preparing is not very desirable, therefore can influence the precision of mensuration.
Show according to the study, it is to be kernel with the polystyrene that there is the polystyrene latex of carboxyl modified on the surface, its surperficial carboxyl can pass through chemical method coupled antigen or coupling antibody, this latex is suitable for use in the latex enhancing immune tuurbidimetry, but, but rarely have report about preparing the polystyrene latex that the surface has than high-carboxyl-content and uniform particle diameter.
Summary of the invention
The present invention is directed to the above-mentioned deficiency of prior art, provide a kind of emulsifier-free residual, the latex particle uniform particle diameter is measured the preparation method that precision height and surface have the polystyrene latex of the surface band carboxyl that is used for immunoassay technology that carboxyl can be by chemical method coupled antigen or coupling antibody.
In order to solve the problems of the technologies described above, the present invention is achieved through the following technical solutions: the preparation method of the polystyrene latex of a kind of surface band carboxyl that is used for immunoassay technology of the present invention, and preparation process comprises:
(1) vinylbenzene (St), methacrylic acid (MAA), water are stirred 10~30min after joining reaction vessel; Above-mentioned methacrylic acid and cinnamic mol ratio are 0.1~0.2: 1;
(2) after the reaction mixture of step (1) gained stirs and finishes, in this reaction mixture, feed the nitrogen (N of 30min 2), in order to remove the oxygen in the reaction mixture, and then in this reaction mixture, add Potassium Persulphate (KPS) aqueous solution and sodium-chlor (NaCl) aqueous solution, make that the concentration of sodium-chlor in the reaction mixture that adds the back gained is that the concentration range of 0.064mol/L, Potassium Persulphate is 7.00 * 10 -4~25.0 * 10 -3Mol/L, and then in the reaction mixture of this moment, continue to feed the N of 20min 2
(3) reaction mixture of step (2) gained stops to feed nitrogen after nitrogen feeds 20min, reaction mixture is warming up to 60~80 ℃ again, reaction 12~24h;
(4) reaction is finished postcooling to room temperature, and obtaining the present invention surface is the polystyrene latex of surface band carboxyl through carboxy-modified polystyrene latex.
The present invention is a kind of to be used for the preparation method of polystyrene latex of the surface band carboxyl of immunoassay technology, preparation process also comprises: with the polystyrene latex of the surface band carboxyl of step (4) gained with deionized water repeatedly centrifugal settling clean, till the specific conductivity of the centrifugal supernatant liquor that goes out and deionized water similar.
The grain diameter of the polystyrene latex of the surface band carboxyl of the present invention's preparation is 100~500nm.
The water that adds in the above-mentioned steps (1) and step (2) preparation persulfate aqueous solution and the used water of sodium chloride aqueous solution, the total amount of two portions institute water will make that the concentration of the added methacrylic acid of step (1) in this total water amount is 0.0819~0.174mol/L.
Sodium chloride electrolysis matter:
Reaction system of the present invention is the soap-free polymerization system, and for the soap-free polymerization system, because used initiator is a Potassium Persulphate, the sulfate radical after the initiator division is adsorbed on around the superpolymer, because the Coulomb repulsion effect keeps the stable of system.When adding sodium chloride electrolysis matter, along with ionic strength increases, the double-deck attenuation of emulsion particle, electrostatic repulsion forces descends gradually, and it is more and more unstable that system becomes, and makes the initial ion loss of stability and condenses each other, thereby the particle diameter of latex particle becomes big, the bigger polymer latex of formation particle diameter.Cause the system unstability because ionic strength increases, latex particle is also skewness therefore, thereby dispersion coefficient increases.When ionic strength increases to a certain degree, it is less to grain diameter influence, and bigger to the dispersiveness influence.
The methacrylic acid comonomer:
The adding of hydrophilic polymerized monomer (methacrylic acid) makes the polystyrene latex particle grain size obviously reduce.This is because the stabilization that hydrophilic polymerized monomer played is far longer than the hydrophilic radical of initiator, makes that the latex particle that generates is more stable, can not assemble between the particle.
Solvent:
The adding of polarity or non-polar solvent owing to the different solubility to monomer and polymkeric substance, therefore has certain influence to latex particle size and dispersiveness.
Advantage of the present invention and beneficial effect:
1. its surface of polystyrene latex of the present invention's preparation has higher carboxyl-content, it is kernel with the polystyrene that there is the polystyrene latex of carboxyl modified on the surface, its surperficial carboxyl is by chemical method coupled antigen or antibody, therefore, be suitable for utilizing the latex enhancing immune tuurbidimetry, measure effective.
2. the present invention is that preparation process is not used emulsifying agent, therefore, has eliminated in the conventional emulsion polymerization product owing to residual emulsifying agent influences its this unfavorable factor of final use properties by the polystyrene latex of emulsifier-free emulsion polymerization preparation.
3. the polystyrene latex of the present invention's preparation is to adopt the emulsifier-free emulsion polymerization preparation, the latex microsphere specific surface area of preparation is big, size distribution is even, surface cleaning, and some character on surface such as the number of wetting ability, functional group and be distributed in to a certain extent and can control, therefore the polystyrene latex that utilizes the present invention to prepare satisfies among the LETIA requirement to latex aspect homogeneity and carboxyl-content.
Description of drawings
The polystyrene latex grain graininess nanometer of Fig. 1 embodiment of the invention 1 gained is analyzed.
The polystyrene latex particle transmission electron microscope figure (TEM) of Fig. 2 embodiment of the invention 1 gained.
Embodiment
Further describe the present invention below in conjunction with specific embodiment, but the present invention is not limited in following embodiment.
Embodiment 1
(1) after being joined there-necked flask, vinylbenzene, methacrylic acid, water stirs 10min; Above-mentioned vinylbenzene is that 0.041mol, methacrylic acid are 8.7mmol, water 10ml;
(2) after the reaction system of step (1) gained stirs and finishes, in reaction system, feed the N of 30min 2Potassium Persulphate that then will about 0.066g and the sodium-chlor of about 0.207g join in the water of 40ml and are configured to the aqueous solution, the aqueous solution of above-mentioned configuration is joined in the reaction system, and the concentration that adds NaCl in the afterreaction system is that the concentration of 0.064mol/L, KPS is 4.38 * 10 -3(the total water amount is that added water of step (1) and step (2) dispose sodium-chlor and the used water of Potassium Persulphate is 50ml to mol/L, configuration NaCl and the used water of the KPS aqueous solution just guarantee to be dissolved in NaCl and KPS in the water fully, because if NaCl and KPS are joined words in the reaction system with solid-state form, there is vinylbenzene to exist, differs and guarantee the solvability of NaCl and KPS surely; As long as determine to add after the NaCl and the KPS aqueous solution, the concentration of NaCl and KPS is that above-mentioned concentration gets final product in the whole reaction system, and embodiment 2~5 is also together), and then continuation feeds the N of 20min 2, in whole reaction system, the total amount of the water that is added is 50ml, the concentration of the added methacrylic acid of step (1) this moment in the total water amount is 0.174mol/L;
The amount of the water that step (1) adds is less than total water amount 50ml, as long as remainder water guarantees NaCl and KPS that complete dissolving step (2) needs adding, therefore, the amount of the water that each step (1) adds and the NaCl and the used water yield of KPS of dissolving step (2) all can be unfixed or fixed, the total water amount is 50ml, and embodiment 2~5 also together.
(3) after nitrogen feeds and to finish, the reaction system of step (2) gained is warming up to 70 ℃, reacted 17.5 hours;
(4) reaction is finished postcooling and obtain the surface through carboxy-modified polystyrene latex after room temperature.The latex that makes with deionized water repeatedly centrifugal settling clean, till the specific conductivity of the centrifugal supernatant liquor that goes out and deionized water similar, dilute with water is preserved with emulsion form then.
Utilize granularity nanometer analyser and transmission electron microscope (TEM), the latex particle size that makes is detected.Utilize the back conductance volumetry that the carboxyl-content on latex surface is measured, the carboxyl-content that records is 2.60mmol/g.
This latex particle size is about 120nm as can be seen by Fig. 1 and Fig. 2, and single dispersion index is 0.017, smooth surface, and form is regular, and uniform particle diameter has monodispersity preferably.
Embodiment 2
(1) after being joined there-necked flask, vinylbenzene, methacrylic acid, water stirs 10min; Above-mentioned vinylbenzene is that 0.041mol, methacrylic acid are 8.7mmol, water 10ml;
(2) after the reaction system of step (1) gained stirs and finishes, in reaction system, feed the N of 30min 2Potassium Persulphate that then will about 0.117g and the sodium-chlor of about 0.207g join in the water of 40ml and are configured to the aqueous solution, the aqueous solution of above-mentioned configuration is joined in the reaction system, and the concentration that adds NaCl in the afterreaction system is that the concentration of 0.064mol/L, KPS is 7.88 * 10 -3Mol/L, and then continuation feeds the N of 20min 2, in whole reaction system, the total amount of the water that is added is 50ml, the concentration of the added methacrylic acid of step (1) this moment in the total water amount is 0.174mol/L;
(3) after nitrogen feeds and to finish, the reaction system of step (2) gained is warming up to 70 ℃, reacted 17.5 hours;
(4) reaction is finished postcooling and obtain the surface through carboxy-modified polystyrene latex after room temperature.The latex that makes with deionized water repeatedly centrifugal settling clean, till the specific conductivity of the centrifugal supernatant liquor that goes out and deionized water similar, dilute with water is preserved with emulsion form then.
Recording this latex particle size by the analysis of granularity nanometer is 163.9nm, and single dispersion index is 0.021, has monodispersity preferably.
Embodiment 3
(1) after being joined there-necked flask, vinylbenzene, methacrylic acid, water stirs 10min; Above-mentioned vinylbenzene is 0.041mol, methacrylic acid 8.7mmol, water 10ml;
(2) after the reaction system of step (1) gained stirs and finishes, in reaction system, feed the N of 30min 2Potassium Persulphate that then will about 0.12g and the sodium-chlor of about 0.207g join in the water of 40ml and are configured to the aqueous solution, the aqueous solution of above-mentioned configuration is joined in the reaction system, and the concentration that adds NaCl in the afterreaction system is that the concentration of 0.064mol/L, KPS is 8.14 * 10 -3Mol/L, and then continuation feeds the N of 20min 2, in whole reaction system, the amount of the water of adding is 50mL, the concentration of the added methacrylic acid of step (1) this moment in the total water amount is 0.174mol/L;
(3) after nitrogen feeds and to finish, the reaction system of step (2) gained is warming up to 70 ℃, reacted 17.5 hours;
(4) reaction is finished postcooling and obtain the surface through carboxy-modified polystyrene latex after room temperature.The latex that makes with deionized water repeatedly centrifugal settling clean, till the specific conductivity of the centrifugal supernatant liquor that goes out and deionized water similar, dilute with water is preserved with emulsion form then.
Recording this latex particle size by the analysis of granularity nanometer is 176.9nm, and single dispersion index is 0.054, has monodispersity preferably.
Embodiment 4
(1) after being joined there-necked flask, vinylbenzene, methacrylic acid, water stirs 10min; Above-mentioned vinylbenzene is that 0.041mol, methacrylic acid are 7.6mmol, water 10ml;
(2) after the reaction system of step (1) gained stirs and finishes, in reaction system, feed the N of 30min 2Potassium Persulphate that then will about 0.118g and the sodium-chlor of about 0.207g join in the water of 40ml and are configured to the aqueous solution, the aqueous solution of above-mentioned configuration is joined in the reaction system, and the concentration that adds NaCl in the afterreaction system is that the concentration of 0.064mol/L, KPS is 7.88 * 10 -3Mol/L, and then continuation feeds the N of 20min 2, in whole reaction system, the amount of the water of adding is 50mL, the concentration of the added methacrylic acid of step (1) this moment in the total water amount is 0.152mol/L;
(3) after nitrogen feeds and to finish, the reaction system of step (2) gained is warming up to 70 ℃, reacted 17.5 hours;
(4) reaction is finished postcooling and obtain the surface through carboxy-modified polystyrene latex after room temperature.The latex that makes with deionized water repeatedly centrifugal settling clean, till the specific conductivity of the centrifugal supernatant liquor that goes out and deionized water similar, dilute with water is preserved with emulsion form then.
Recording this latex particle size by the analysis of granularity nanometer is 204.0nm, and single dispersion index is 0.025, has monodispersity preferably.
Embodiment 5
(1) after being joined there-necked flask, vinylbenzene, methacrylic acid, water stirs 10min; Above-mentioned vinylbenzene is that 0.041mol, methacrylic acid are 7mmol, water 12ml;
(2) after the reaction system of step (1) gained stirs and finishes, in reaction system, feed the N of 30min 2Potassium Persulphate that then will about 0.118g and the sodium-chlor of about 0.207g join in the water of 38ml and are configured to the aqueous solution, the aqueous solution of above-mentioned configuration is joined in the reaction system, and the concentration that adds NaCl in the afterreaction system is that the concentration range of 0.064mol/L, KPS is 7.88 * 10 -3Mol/L, and then continuation feeds the N of 20min 2, in whole reaction system, the amount of the water of adding is 50mL, the concentration of the added methacrylic acid of step (1) this moment in the total water amount is 0.14mol/L;
(3) after nitrogen feeds and to finish, the reaction system of step (2) gained is warming up to 70 ℃, reacted 17.5 hours;
(4) reaction is finished postcooling and obtain the surface through carboxy-modified polystyrene latex after room temperature.The latex that makes with deionized water repeatedly centrifugal settling clean, till the specific conductivity of the centrifugal supernatant liquor that goes out and deionized water similar, dilute with water is preserved with emulsion form then.
Recording this latex particle size by the analysis of granularity nanometer is 251.0nm, and single dispersion index is 0.051, has monodispersity preferably.

Claims (2)

1. the preparation method of the polystyrene latex of a surface band carboxyl that is used for immunoassay technology, it is characterized in that: preparation process comprises:
(1) after being joined reaction vessel, vinylbenzene, methacrylic acid, water stirs 10~30min; Above-mentioned methacrylic acid and cinnamic mol ratio are 0.1~0.2: 1;
(2) after the reaction mixture of step (1) gained stirs and finishes, in this reaction mixture, feed the nitrogen of 30min, in order to remove the air in the reaction mixture, and then in this reaction mixture, add persulfate aqueous solution and sodium chloride aqueous solution, make that the concentration of NaCl in the reaction mixture that adds the back gained is that the concentration range of 0.064mol/L, Potassium Persulphate is 7.00 * 10 -4~25.0 * 10 -3Mol/L, and then in the reaction mixture of this moment, continue to feed the nitrogen of 20min;
(3) reaction mixture of step (2) gained stops to feed nitrogen after nitrogen feeds 20min, again reaction mixture at this moment is warming up to 60~80 ℃, reaction 12~24h;
(4) reaction is finished postcooling to room temperature, obtains the surface promptly has carboxyl through carboxy-modified polystyrene latex polystyrene latex;
Preparation persulfate aqueous solution and the used water of sodium chloride aqueous solution in water that adds in the described step (1) and the step (2), the total amount of two portions institute water will make that the concentration of the added methacrylic acid of step (1) in this total water amount is 0.0819~0.174mol/L.
2. the preparation method of the polystyrene latex of the surface band carboxyl that is used for immunoassay technology according to claim 1 is characterized in that: the grain diameter of the polystyrene latex of surface band carboxyl is 100~500nm.
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CN107177019B (en) * 2016-03-11 2019-09-27 中国石油化工股份有限公司 A kind of polystyrene emulsion and its preparation method and application
CN106432572B (en) * 2016-08-10 2019-05-24 太原工业学院 A kind of polymer material and its preparation method and application with acid-base discoloration instruction function
CN107894509B (en) * 2017-10-20 2019-06-28 柏荣诊断产品(上海)有限公司 A method of improving latex immunoturbidimetry antigen excess and the range of linearity
CN110846335B (en) * 2018-12-22 2023-02-28 波音特生物科技(南京)有限公司 Preparation method of antibody kit capable of detecting highly pathogenic strains of helicobacter pylori
CN113150200A (en) * 2021-03-16 2021-07-23 苏州为度生物技术有限公司 Preparation method and application of carboxyl latex microspheres

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