CN106674401A - Preparation method of polystyrene latex with carboxyl on surface for immunodetection technique - Google Patents
Preparation method of polystyrene latex with carboxyl on surface for immunodetection technique Download PDFInfo
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- CN106674401A CN106674401A CN201510754396.3A CN201510754396A CN106674401A CN 106674401 A CN106674401 A CN 106674401A CN 201510754396 A CN201510754396 A CN 201510754396A CN 106674401 A CN106674401 A CN 106674401A
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Abstract
The invention discloses a preparation method of polystyrene latex with carboxyl on a surface for an immunodetection technique. The preparation method comprises the following preparation steps of adding styrene, methacrylic acid and water into a three-mouth flask, and then agitating an obtained mixture for 10min to 30min, wherein the mole ratio of the methacrylic acid to the styrene is (0.1 to 0.2) to 1; after the agitation is terminated, introducing N2 into a reaction system, then adding the aqueous solution of potassium persulfate and sodium chloride into the reaction system, so as to enable the concentration of the sodium chloride in the reaction system to be 0.064mol/L and the concentration range of the potassium persulfate to be 7.00*10<-4>mol/L to 25.0*10<-3>mol/L, and then continuously introducing the N2 into the reaction system again; raising a temperature to 60 to 80 DEG C, and carrying out a reaction for 12h to 24h; after the reaction is completed, cooling the reaction system to a room temperature to subsequently obtain a target product of the preparation method. The latex prepared by the preparation method is uniform in particle size, good in monodispersity and high in stability; meanwhile, the latex has the higher content of the carbonyl on the surface, and is suitably used for a latex enhanced immunoturbidimetry.
Description
Technical field
The present invention relates to a kind of preparation method of the polystyrene latex of the surface band carboxyl for immunoassay technology.
Background technology
In field of immunodetection generally, first antigen-antibody reaction is formed into immune complex, then quantitative determined with nephelometry, but form antigen-antibody complex needs the long period.Latex enhancing immune turbidimetry (abbreviation LETIA) is a kind of new immunoassay technology that in recent years field of immunodetection occurs, LETIA does not need specific apparatus, automatically, semi-automatic biochemical analyzer can be determined, suitably in vast basic medical treatment unit popularization and application.Being applied to many analyte quantitative determinations at present has as many as tens of kinds, such as ASO, RF, CRP, transferrins, toxoplasm, rubella, β2- microglobulin and drug surveillance etc..
LETIA its principle is the present latex particulate (immune latex) for being coated with antigen or antibody, occur after immune response with corresponding antibodies in sample or antigen, form agglutinating particle, the formation of agglutinator makes reaction hybrid system form certain turbidity, the degree that turbidity increases is proportional with checking matter concentration in sample, turbidimetric analysis turbidimetry is carried out under certain wavelength, you can measure the content of checking matter in sample.Therefore, the key that suitable latex (immune latex) is latex enhancing immune turbidimetric analysis turbidimetry how is prepared.
At present, after the completion of conventional latex preparation process, will use emulsifying agent as auxiliary material more, but prepared by latex, wherein a certain amount of emulsifying agent is often remained, and the presence of emulsifying agent can affect the final performance of latex.
Separately, the wavelength of latex enhancing immune turbidimetric analysis turbidimetry is relevant with latex particle size size used, and the uniform particle diameter of latex particle prepared by prior art is less desirable, therefore can affect the precision for determining.
Show according to the study, the polystyrene latex that there is carboxyl modified on surface is with polystyrene as kernel, the carboxyl on its surface can be by chemical method coupled antigen or coupled antibody, the latex is suitable for use in latex enhancing immune turbidimetry, but, with regard to preparing, surface has compared with high-carboxyl-content and the polystyrene latex of uniform particle diameter is but rarely reported.
The content of the invention
Above-mentioned deficiency of the present invention for prior art, a kind of emulsifier-free residual is provided, latex particle uniform particle diameter, measurement accuracy is high and surface can be by chemical method coupled antigen or the preparation method of the polystyrene latex of the surface band carboxyl for immunoassay technology of coupled antibody with carboxyl.
In order to solve above-mentioned technical problem, the present invention is achieved through the following technical solutions：A kind of preparation method of the polystyrene latex of surface band carboxyl for immunoassay technology of the present invention, preparation process includes：
(1) styrene (St), methacrylic acid (MAA), water are added into 10～30min of stirring after reaction vessel；Above-mentioned methacrylic acid is 0.1～0.2 with cinnamic mol ratio: 1；
(2) after the reactant mixture stirring obtained by step (1) terminates, the nitrogen (N of 30min is passed through in the reactant mixture2), oxygen in remove reactant mixture, then potassium peroxydisulfate (KPS) aqueous solution and sodium chloride (NaCl) aqueous solution are added in the reactant mixture again so that after adding in the reactant mixture of gained the concentration of sodium chloride be 0.064mol/L, the concentration range of potassium peroxydisulfate be 7.00 × 10-4～25.0 × 10-3Mol/L, then continues in reactant mixture now again to be passed through the N of 20min2；
(3) after nitrogen is passed through 20min, stopping is passed through nitrogen to the reactant mixture obtained by step (2), then reactant mixture is warming up into 60～80 DEG C, reacts 12～24h；
(4) room temperature is cooled to after the completion of reacting, surface of the present invention is obtained through the carboxy-modified polystyrene latex i.e. polystyrene latex of surface band carboxyl.
A kind of preparation method of the polystyrene latex of the surface band carboxyl for immunoassay technology of the present invention, preparation process also includes：By the polystyrene latex deionized water of the surface band carboxyl obtained by step (4) multiple centrifugal sedimentation cleaning, until the electrical conductivity of the supernatant being centrifuged out to till the similar of deionized water.
The grain diameter of the polystyrene latex of surface band carboxyl prepared by the present invention is 100～500nm.
Water used by water and step (2) the preparation persulfate aqueous solution of addition in above-mentioned steps (1) and sodium-chloride water solution, it is 0.0819～0.174mol/L that the total amount of water used by two parts will cause concentration of the methacrylic acid added by step (1) in the total water consumption.
Sodium chloride electrolysis matter：
The reaction system of the present invention is soap-free polymerization system, and for soap-free polymerization system, because initiator used is potassium peroxydisulfate, the sulfate radical after initiator division adsorbs around high polymer, because electrostatic repulsion keeps stablizing for system.When sodium chloride electrolysis matter is added, as ionic strength increases, emulsion particle bilayer is thinning, electrostatic repulsion forces are gradually reduced, and system becomes more and more unstable so that initial ion loss of stability and condense each other, thus the particle diameter of latex particle becomes big, the larger polymer emulsion of particle diameter is formed.Because ionic strength increase causes system unstability, latex particle therefore also skewness, so as to coefficient of dispersion increase.When ionic strength increases to a certain extent, it is less to grain diameter influence, and affects larger to dispersiveness.
Methacrylic acid comonomers：
The addition of hydrophilic co-monomer (methacrylic acid) so that the particle diameter of polystyrene latex particles is obviously reduced.This is because the stabilization that hydrophilic co-monomer is played is far longer than the hydrophilic radical of initiator so that the latex particle of generation is more stable, will not assemble between particle.
Solvent：
The addition of polarity or non-polar solven, due to the different solubility to monomer and polymer, therefore has certain impact to latex particle size and dispersiveness.
The advantages of the present invention：
1. its surface of polystyrene latex that prepared by the present invention has higher carboxyl-content, there is the polystyrene latex of carboxyl modified on surface with polystyrene as kernel, and the carboxyl on its surface passes through chemical method coupled antigen or antibody, therefore, it is suitable to, using latex enhancing immune turbidimetry, determine effect good.
2. the present invention is the polystyrene latex prepared by emulsifier-free emulsion polymerization, and preparation process uses emulsifying agent, therefore, to eliminate in conventional emulsion polymerization product and affect its end-use properties this unfavorable factor due to residual emulsifying agents.
3. the polystyrene latex that prepared by the present invention is prepared using emulsifier-free emulsion polymerization, the latex microsphere specific surface area of preparation is big, even particle size distribution, clean surface, and the properties on the surface such as number of hydrophily, functional group and distribution can be controlled to a certain extent, meet the requirement in LETIA to latex in terms of homogeneity and carboxyl-content hence with the polystyrene latex that the present invention is prepared.
Description of the drawings
The polystyrene latex particles granularity nanoanalysis of the gained of Fig. 1 embodiment of the present invention 1.
The polystyrene latex particles transmission electron microscope figure (TEM) of the gained of Fig. 2 embodiment of the present invention 1.
Specific embodiment
The present invention is further described with reference to specific embodiment, but the present invention is not limited in following embodiments.
Embodiment 1
(1) styrene, methacrylic acid, water are added to after there-necked flask and stir 10min；Above-mentioned styrene is 0.041mol, methacrylic acid is 8.7mmol, water 10ml；
(2) after the reaction system stirring obtained by step (1) terminates, the N of 30min is passed through in reaction system2Then the sodium chloride of the potassium peroxydisulfate of about 0.066g and about 0.207g is added in the water of 40ml and is configured to the aqueous solution, the aqueous solution of above-mentioned configuration is added in reaction system, the concentration for adding NaCl in rear reaction system is 4.38 × 10 for the concentration of 0.064mol/L, KPS-3Mol/L (total water consumption be the water added by step (1) and step (2) configuration sodium chloride and potassium peroxydisulfate used by water be 50ml, water used by the configuration NaCl and KPS aqueous solution is only to ensure that and NaCl and KPS can be dissolved completely in water, if because NaCl and KPS are added in reaction system in the form of solid-state, with the presence of styrene, the dissolubility of NaCl and KPS is not necessarily can ensure that；Add after NaCl the and KPS aqueous solution as long as determining, the concentration of NaCl and KPS is above-mentioned concentration in whole reaction system, embodiment 2～5 is also same), then it is further continued for being passed through the N of 20min2, in whole reaction system, the total amount of the water for being added is 50ml, and now concentration of the methacrylic acid added by step (1) in total water consumption is 0.174mol/L；
The amount of the water that step (1) is added is less than total water consumption 50ml, as long as remaining water guarantees to be completely dissolved NaCl and KPS that step (2) needs to add, therefore, every time the water yield used by the amount of water and the NaCl and KPS of dissolving step (2) of step (1) addition may each be unfixed or fixed, total water consumption is 50ml, and embodiment 2～5 is also same.
(3) after the completion of nitrogen is passed through, the reaction system obtained by step (2) is warming up into 70 DEG C, is reacted 17.5 hours；
(4) it is cooled to after room temperature after the completion of reacting and obtains surface through carboxy-modified polystyrene latex.The multiple centrifugal sedimentation cleaning of obtained latex deionized water, until the electrical conductivity of the supernatant being centrifuged out to till the similar of deionized water, then dilute with water, preserves in the form of an emulsion.
Using granularity nanoanalysis instrument and transmission electron microscope (TEM), obtained latex particle size is detected.The carboxyl-content of latex surface is determined using back conductance titration, the carboxyl-content for measuring is 2.60mmol/g.
The latex particle size can be seen that in 120nm or so by Fig. 1 and Fig. 2, single dispersing index is 0.017, and surface is smooth, and form is regular, uniform particle diameter, with preferable monodispersity.
Embodiment 2
(1) styrene, methacrylic acid, water are added to after there-necked flask and stir 10min；Above-mentioned styrene is 0.041mol, methacrylic acid is 8.7mmol, water 10ml；
(2) after the reaction system stirring obtained by step (1) terminates, the N of 30min is passed through in reaction system2Then the sodium chloride of the potassium peroxydisulfate of about 0.117g and about 0.207g is added in the water of 40ml and is configured to the aqueous solution, the aqueous solution of above-mentioned configuration is added in reaction system, the concentration for adding NaCl in rear reaction system is 7.88 × 10 for the concentration of 0.064mol/L, KPS-3Mol/L, is then further continued for being passed through the N of 20min2, in whole reaction system, the total amount of the water for being added is 50ml, and now concentration of the methacrylic acid added by step (1) in total water consumption is 0.174mol/L；
(3) after the completion of nitrogen is passed through, the reaction system obtained by step (2) is warming up into 70 DEG C, is reacted 17.5 hours；
(4) it is cooled to after room temperature after the completion of reacting and obtains surface through carboxy-modified polystyrene latex.The multiple centrifugal sedimentation cleaning of obtained latex deionized water, until the electrical conductivity of the supernatant being centrifuged out to till the similar of deionized water, then dilute with water, preserves in the form of an emulsion.
It is 163.9nm to measure the latex particle size by granularity nanoanalysis, and single dispersing index is 0.021, with preferable monodispersity.
Embodiment 3
(1) styrene, methacrylic acid, water are added to after there-necked flask and stir 10min；Above-mentioned styrene be 0.041mol, methacrylic acid 8.7mmol, water 10ml；
(2) after the reaction system stirring obtained by step (1) terminates, the N of 30min is passed through in reaction system2Then the sodium chloride of the potassium peroxydisulfate of about 0.12g and about 0.207g is added in the water of 40ml and is configured to the aqueous solution, the aqueous solution of above-mentioned configuration is added in reaction system, the concentration for adding NaCl in rear reaction system is 8.14 × 10 for the concentration of 0.064mol/L, KPS-3Mol/L, is then further continued for being passed through the N of 20min2, in whole reaction system, the amount of the water of addition is 50mL, and now concentration of the methacrylic acid added by step (1) in total water consumption is 0.174mol/L；
(3) after the completion of nitrogen is passed through, the reaction system obtained by step (2) is warming up into 70 DEG C, is reacted 17.5 hours；
(4) it is cooled to after room temperature after the completion of reacting and obtains surface through carboxy-modified polystyrene latex.The multiple centrifugal sedimentation cleaning of obtained latex deionized water, until the electrical conductivity of the supernatant being centrifuged out to till the similar of deionized water, then dilute with water, preserves in the form of an emulsion.
It is 176.9nm to measure the latex particle size by granularity nanoanalysis, and single dispersing index is 0.054, with preferable monodispersity.
Embodiment 4
(1) styrene, methacrylic acid, water are added to after there-necked flask and stir 10min；Above-mentioned styrene is 0.041mol, methacrylic acid is 7.6mmol, water 10ml；
(2) after the reaction system stirring obtained by step (1) terminates, the N of 30min is passed through in reaction system2Then the sodium chloride of the potassium peroxydisulfate of about 0.118g and about 0.207g is added in the water of 40ml and is configured to the aqueous solution, the aqueous solution of above-mentioned configuration is added in reaction system, the concentration for adding NaCl in rear reaction system is 7.88 × 10 for the concentration of 0.064mol/L, KPS-3Mol/L, is then further continued for being passed through the N of 20min2, in whole reaction system, the amount of the water of addition is 50mL, and now concentration of the methacrylic acid added by step (1) in total water consumption is 0.152mol/L；
(3) after the completion of nitrogen is passed through, the reaction system obtained by step (2) is warming up into 70 DEG C, is reacted 17.5 hours；
(4) it is cooled to after room temperature after the completion of reacting and obtains surface through carboxy-modified polystyrene latex.The multiple centrifugal sedimentation cleaning of obtained latex deionized water, until the electrical conductivity of the supernatant being centrifuged out to till the similar of deionized water, then dilute with water, preserves in the form of an emulsion.
It is 204.0nm to measure the latex particle size by granularity nanoanalysis, and single dispersing index is 0.025, with preferable monodispersity.
Embodiment 5
(1) styrene, methacrylic acid, water are added to after there-necked flask and stir 10min；Above-mentioned styrene is 0.041mol, methacrylic acid is 7mmol, water 12ml；
(2) after the reaction system stirring obtained by step (1) terminates, the N of 30min is passed through in reaction system2Then the sodium chloride of the potassium peroxydisulfate of about 0.118g and about 0.207g is added in the water of 38ml and is configured to the aqueous solution, the aqueous solution of above-mentioned configuration is added in reaction system, the concentration for adding NaCl in rear reaction system is 7.88 × 10 for the concentration range of 0.064mol/L, KPS-3Mol/L, is then further continued for being passed through the N of 20min2, in whole reaction system, the amount of the water of addition is 50mL, and now concentration of the methacrylic acid added by step (1) in total water consumption is 0.14mol/L；
(3) after the completion of nitrogen is passed through, the reaction system obtained by step (2) is warming up into 70 DEG C, is reacted 17.5 hours；
(4) it is cooled to after room temperature after the completion of reacting and obtains surface through carboxy-modified polystyrene latex.The multiple centrifugal sedimentation cleaning of obtained latex deionized water, until the electrical conductivity of the supernatant being centrifuged out to till the similar of deionized water, then dilute with water, preserves in the form of an emulsion.
It is 251.0nm to measure the latex particle size by granularity nanoanalysis, and single dispersing index is 0.051, with preferable monodispersity.
Claims (4)
1. a kind of preparation method of the polystyrene latex of the surface band carboxyl for immunoassay technology, it is characterised in that：Preparation process includes：
(1) styrene, methacrylic acid, water are added into 10～30min of stirring after reaction vessel；Above-mentioned methacrylic acid is 0.1～0.2 with cinnamic mol ratio: 1；
(2) after the reactant mixture stirring obtained by step (1) terminates, the nitrogen of 30min is passed through in the reactant mixture, air in remove reactant mixture, then persulfate aqueous solution and sodium-chloride water solution are added in the reactant mixture again so that after adding in the reactant mixture of gained the concentration of NaCl be 0.064mol/L, the concentration range of potassium peroxydisulfate be 7.00 × 10-4～25.0 × 10-3Mol/L, then continues in reactant mixture now again to be passed through the nitrogen of 20min；
(3) after nitrogen is passed through 20min, stopping is passed through nitrogen to the reactant mixture obtained by step (2), then reactant mixture now is warming up into 60～80 DEG C, reacts 12～24h；
(4) room temperature is cooled to after the completion of reacting, surface of the present invention is obtained through the carboxy-modified polystyrene latex i.e. polystyrene latex with carboxyl.
2. the preparation method of the polystyrene latex of the surface band carboxyl for immunoassay technology according to claim 1, it is characterised in that：Preparation process also includes：By the polystyrene latex deionized water of the surface band carboxyl obtained by above-mentioned steps (4) multiple centrifugal sedimentation cleaning, until the electrical conductivity of the supernatant being centrifuged out to till the similar of deionized water.
3. the preparation method of the polystyrene latex of the surface band carboxyl for immunoassay technology according to claim 1 and 2, it is characterised in that：The grain diameter of the polystyrene latex of surface band carboxyl is 100～500nm.
4. the preparation method of the polystyrene latex of the surface band carboxyl for immunoassay technology according to claim 1, it is characterised in that：Water used by the water and the middle preparation persulfate aqueous solution of step (2) of the middle addition of the step (1) and sodium-chloride water solution, it is 0.0819～0.174mol/L that the total amount of water used by two parts will cause concentration of the methacrylic acid added by step (1) in the total water consumption.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108383931A (en) * | 2018-01-12 | 2018-08-10 | 武汉生之源生物科技股份有限公司 | A kind of preparation method of chloromethylated polystyrene latex |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108383931A (en) * | 2018-01-12 | 2018-08-10 | 武汉生之源生物科技股份有限公司 | A kind of preparation method of chloromethylated polystyrene latex |
| CN108383931B (en) * | 2018-01-12 | 2020-06-19 | 武汉生之源生物科技股份有限公司 | Preparation method of chloromethylated polystyrene latex |
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