CN113150352A - Preparation method and application of surface-activated fluorescent latex microspheres - Google Patents

Preparation method and application of surface-activated fluorescent latex microspheres Download PDF

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CN113150352A
CN113150352A CN202110289748.8A CN202110289748A CN113150352A CN 113150352 A CN113150352 A CN 113150352A CN 202110289748 A CN202110289748 A CN 202110289748A CN 113150352 A CN113150352 A CN 113150352A
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fluorescent latex
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stirring
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宋孟杰
刘照关
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Suzhou Vdo Biotech Co ltd
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Abstract

The invention relates to a preparation method of a surface activated fluorescent latex microsphere, which comprises the following steps: dispersing the fluorescent latex microspheres with the surface modified with hydroxyl groups, adding a catalyst, stirring until the catalyst is dissolved, adding epoxy chloropropane, and stirring; adding alkali, stirring and cleaning to obtain the surface activated fluorescent latex microspheres. The invention obtains the fluorescent latex microsphere with the surface modified by epoxy groups by modifying hydroxyl groups, the microsphere with the activated surface can be combined with an antibody by one step, the coupling process is simplified, and higher reaction activity is shown in antibody coupling.

Description

Preparation method and application of surface-activated fluorescent latex microspheres
Technical Field
The invention relates to the technical field of high polymer materials, in particular to a preparation method and application of a surface-activated fluorescent latex microsphere.
Background
The latex microspheres are nano-scale spherical marking materials synthesized by high polymer materials, can be classified into carboxylated, aminated and sulfhydrylated latex microspheres according to different surface functional groups, and can wrap dyes with different colors, so that the colors of the latex microspheres are rich, and the requirements of various experiments can be met. Because of the stability problem, before the fluorescent latex microsphere is used, the surface of the microsphere is not activated and has no adsorption or chemical adhesive capacity, and when the fluorescent latex microsphere is used, the fluorescent latex microsphere can be covalently combined with an antibody or an antigen by virtue of abundant functional groups (such as carboxyl, amino and the like) on the surface of the microsphere through activation treatment, so as to capture a corresponding object to be detected, and then the quantitative detection of corresponding biomolecules is realized through the strength of a fluorescent signal.
The coupling method commonly used for covalently bonding the microspheres and the antigen/antibody is EDC/NHS method. Firstly, activating carboxyl functional groups on the microspheres by using water-soluble carbodiimide EDC to generate an active ester intermediate, wherein the active ester intermediate can be directly and efficiently combined with amino-containing biomolecules, so that the biomolecules are coupled to the surfaces of the microspheres. However, the active ester intermediate formed by EDC reaction has poor stability and is easy to hydrolyze, and another more stable active ester intermediate can be formed by introducing N-hydroxysuccinimide (NHS) and can perform substitution reaction with amino to form amido bond, thereby greatly improving the binding efficiency of biomolecules. However, in the activation coupling process, the reagent needs to be eluted after the activation is completed, the general flow includes activation, centrifugation, washing, dispersion, protein addition for combination, and reagent preparation, and many unstable situations still exist in the process, such as microsphere agglomeration, poor activation effect, and low coupling efficiency, and how to more efficiently and stably combine microspheres and biomolecules becomes a key problem to be solved urgently in the biological application of latex microspheres.
CN1246351C discloses an epoxy functional crosslinking core-shell structure nano polymer microsphere, which is obtained by emulsion or suspension polymerization in the presence of an emulsifier and an initiator, wherein the emulsion or suspension polymerization takes water as a medium, and the polymer microsphere has a core-shell structure, a nanoscale and epoxy functional groups on the surface; CN103386282B adopts silane coupling agent with epoxy group to obtain magnetic microsphere with activated surface epoxy group, but the construction mode is more complicated.
Disclosure of Invention
In order to solve the technical problems, the fluorescent latex microspheres with epoxy group modified surfaces are obtained by modifying hydroxyl groups, the microspheres with activated surfaces can be combined with antibodies in one step, and the coupling process is simplified, so that the problems of microsphere agglomeration, poor activation effect and low coupling efficiency in the traditional activation coupling process are solved.
The invention relates to a preparation method of a surface activated fluorescent latex microsphere, which comprises the following steps:
(1) dispersing the fluorescent latex microspheres with the surface modified with hydroxyl groups, adding a catalyst, stirring until the catalyst is dissolved, adding epoxy chloropropane, and stirring;
(2) and (2) adding alkali into the mixed solution obtained in the step (1), stirring, and cleaning to obtain the fluorescent latex microspheres with activated surfaces.
Under the action of a catalyst, epoxy groups on epoxy chloropropane react with hydroxyl groups on the surface of the microsphere to open the ring, the epoxy chloropropane after the ring opening is grafted to the surface of the hydroxyl microsphere, and then dehydrochlorination and ring closure are carried out under an alkaline condition to obtain the microsphere with the epoxy group surface. The covalent connection of the epoxy group activated microspheres and biomolecules does not need the addition of other activating agents, and meanwhile, the two-step reaction improves the reaction efficiency of the hydroxyl groups on the surfaces of the microspheres and ensures the high yield of the subsequent epoxy groups. The two-step reaction is as follows:
Figure BDA0002978594810000021
further, the mass ratio of the fluorescent latex microspheres with hydroxyl groups modified on the surface, the catalyst, the epichlorohydrin and the alkali is 1:0.01-0.1:0.1-10: 0.1-10.
Further, the mass ratio of the epichlorohydrin to the alkali is 1: 1-1.1. The purpose of this mass ratio is that the epoxy groups obtained by ring closure do not undergo alkaline ring opening in the presence of an excessively high alkaline solution, thereby ensuring the yield of epoxy groups.
Further, in the step (1), the fluorescent latex microspheres are selected from one or more of fluorescent polystyrene microspheres, polymethyl methacrylate microspheres and polyglycidyl methacrylate microspheres, and the size is 100-1000 nm.
Further, in the step (1), the catalyst is a Lewis acid.
Further, the Lewis acid is selected from one or more of aluminum trichloride, boron trifluoride or trifluoromethanesulfonic acid.
Further, in the step (1), after the catalyst is added, the reaction is stirred for 10 to 20 hours at the temperature of between 50 and 90 ℃.
Further, in the step (1), the dripping time of the epichlorohydrin is 0.5-2 h.
Further, in the step (2), the base is selected from one or more of sodium hydroxide, potassium hydroxide and tetramethylammonium hydroxide.
Further, in the step (2), after adding the alkali, stirring and reacting for 2-4h at 40-60 ℃.
Further, in the step (2), the washing is performed by centrifugation using deionized water and absolute ethyl alcohol in sequence.
Further, in the step (2), a drying step is further included after the cleaning.
The invention claims the surface-activated fluorescent latex microspheres prepared by the preparation method.
The invention also claims the application of the surface activated fluorescent latex microspheres prepared by the preparation method in biological coupling.
By the scheme, the invention at least has the following advantages:
(1) according to the invention, the fluorescent latex microspheres with activated surface epoxy groups are obtained by modifying the surface hydroxyl groups of the microspheres with the micromolecule epoxide, so that the process flow of coupling the microspheres with biomolecules is greatly simplified, the repeatability is strong, and the coupling process is more stable and controllable.
(2) The activated microsphere can directly react with ligand molecules such as antibody, protein, amino acid and the like containing amino, sulfydryl and the like.
The foregoing description is only an overview of the technical solutions of the present invention, and in order to make the technical solutions of the present invention more clearly understood and to implement them in accordance with the contents of the description, the following description is made with reference to the preferred embodiments of the present invention and the accompanying detailed drawings.
Drawings
In order that the present disclosure may be more readily and clearly understood, reference is now made to the following detailed description of the embodiments of the present disclosure taken in conjunction with the accompanying drawings, in which
FIG. 1 shows the result of antibody coupling detection of the fluorescent latex microspheres prepared in example 1.
Detailed Description
The present invention is further described below in conjunction with the following figures and specific examples so that those skilled in the art may better understand the present invention and practice it, but the examples are not intended to limit the present invention.
Example 1
Dispersing 1.0g of surface hydroxyl fluorescent latex microspheres (polystyrene material, microsphere size is 100nm) in deionized water, adding 0.1g of aluminum trichloride, stirring to dissolve, weighing 10.0g of epoxy chloropropane, slowly dropwise adding for 1h, stirring at 80 ℃ for reaction for 20h, cooling the reaction liquid to 50 ℃, adding 4.5g of sodium hydroxide, and stirring at 50 ℃ for reaction for 2 h; and cooling the reaction solution to room temperature, sequentially washing with deionized water and absolute ethyl alcohol, and drying to obtain the surface-activated fluorescent latex microspheres.
Example 2
Dispersing 1.0g of surface hydroxyl fluorescent latex microspheres (made of polymethyl methacrylate and with the size of 500nm) in deionized water, adding 0.1g of boron trifluoride into the deionized water, stirring to dissolve the boron trifluoride, weighing 4.0g of epoxy chloropropane, slowly dropwise adding the epoxy chloropropane for 0.5h, stirring at 70 ℃ for reaction for 15h, cooling the reaction liquid to 50 ℃, adding 1.8g of potassium hydroxide into the reaction liquid, and stirring at 50 ℃ for reaction for 3 h; and cooling the reaction solution to room temperature, sequentially washing with deionized water and absolute ethyl alcohol, and drying to obtain the surface-activated fluorescent latex microspheres.
Antibody coupling test:
weighing 100mg of the surface-activated fluorescent latex microsphere powder prepared in example 1, ultrasonically dispersing the powder in 10mL of sodium carbonate buffer (0.1M, pH 10), adding the polyclonal antibody against C-reactive protein (0.1mL, 2mg/mL), fully mixing the powder uniformly, and incubating the mixture at 37 ℃ overnight; adding cysteine, continuing to incubate for 2h, washing with sodium carbonate buffer solution for three times, and finally dispersing in 100mL of sodium carbonate buffer solution to obtain the fluorescent microspheres with the surface coupled with the C-reactive protein polyclonal antibody.
The test paper strip for detecting the C-reactive protein is prepared by taking the obtained fluorescent microspheres with the surface coupled with the polyclonal antibody of the C-reactive protein as a solid phase carrier, a series of C-reactive protein standard substances (0, 0.625, 1.25, 2.5, 5 and 10mg/L) with different concentrations are respectively added into a sample adding area of the prepared test paper strip for detecting the C-reactive protein, a buffer solution is dripped into the sample adding area, a signal value of a T line on the test paper strip is read by a fluorescent quantitative immunoassay analyzer after 10min, and the result is shown in a table 1. The average values of the signals obtained in table 1 were subjected to data processing to obtain a coordinate curve, as shown in fig. 1.
TABLE 1 test results for different concentrations of standards
Figure BDA0002978594810000051
As can be seen from the figure, when the fluorescent latex microsphere prepared by the invention is used for coupling the C-reactive protein antibody, the T-line signal value changes along with the concentration change of the C-reactive protein standard substance to be detected, and the linear correlation coefficient R20.9982, showing that the fluorescent microsphere with the surface coupled with the C-reactive protein polyclonal antibody has good reactivity.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications of the invention may be made without departing from the spirit or scope of the invention.

Claims (10)

1. A preparation method of surface activated fluorescent latex microspheres is characterized by comprising the following steps:
(1) dispersing the fluorescent latex microspheres with the surface modified with hydroxyl groups, adding a catalyst, stirring until the catalyst is dissolved, adding epoxy chloropropane, and stirring;
(2) and (2) adding alkali into the mixed solution obtained in the step (1), stirring, and cleaning to obtain the fluorescent latex microspheres with activated surfaces.
2. The method of claim 1, wherein: the mass ratio of the fluorescent latex microsphere with the surface modified with hydroxyl groups to the catalyst to the epichlorohydrin to the alkali is 1:0.01-0.1:0.1-10: 0.1-10.
3. The method of claim 2, wherein: the mass ratio of the epichlorohydrin to the alkali is 1: 1-1.1.
4. The method of claim 1, wherein: in the step (1), the fluorescent latex microspheres are selected from one or more of fluorescent polystyrene microspheres, polymethyl methacrylate microspheres and polyglycidyl methacrylate microspheres.
5. The method of claim 1, wherein: in the step (1), the catalyst is Lewis acid.
6. The method of claim 1, wherein: in the step (1), after the catalyst is added, stirring and reacting for 10-20h at 50-90 ℃.
7. The method of claim 1, wherein: in step (2), the base is selected from one or more of sodium hydroxide, potassium hydroxide and tetramethylammonium hydroxide.
8. The method of claim 1, wherein: in the step (2), after adding alkali, stirring and reacting for 2-4h at 40-60 ℃.
9. A surface-activated fluorescent latex microsphere prepared by the preparation method of any one of claims 1 to 8.
10. Use of the surface-activated fluorescent latex microspheres of claim 9 in bioconjugation.
CN202110289748.8A 2021-03-16 2021-03-16 Preparation method and application of surface-activated fluorescent latex microspheres Pending CN113150352A (en)

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Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101306352A (en) * 2008-01-30 2008-11-19 上海华明高技术（集团）有限公司 Large particle diameter mono-disperse polymer microspheres containing epoxy group and preparation method thereof
CN101805483A (en) * 2010-02-26 2010-08-18 光景生物科技(苏州)有限公司 Surface functionalization resonance luminous microspheres, kit containing resonance luminous microspheres and application
CN102604305A (en) * 2012-02-21 2012-07-25 苏州纳诺康生物技术有限公司 Nano-grade fluorescent microspheres, preparation method thereof, and purpose thereof as markers used in immunity chromatography method
CN103333299A (en) * 2013-07-12 2013-10-02 西南大学 Glycidyl methacrylate and poly (glycol) methacrylate copolymer microsphere, preparation method and application of copolymer microsphere
CN106380617A (en) * 2016-09-06 2017-02-08 北京华科泰生物技术有限公司 Surface-activated fluorescent latex microspheres as well as preparation method and application thereof
CN106633168A (en) * 2016-10-19 2017-05-10 扬州倍赛德生物科技有限公司 Preparation method of monodisperse and large-particle-diameter polymer microspheres
CN112341663A (en) * 2020-10-28 2021-02-09 苏州纳微科技股份有限公司 ProteinA affinity chromatography medium of PMMA matrix and preparation method and application thereof

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101306352A (en) * 2008-01-30 2008-11-19 上海华明高技术（集团）有限公司 Large particle diameter mono-disperse polymer microspheres containing epoxy group and preparation method thereof
CN101805483A (en) * 2010-02-26 2010-08-18 光景生物科技(苏州)有限公司 Surface functionalization resonance luminous microspheres, kit containing resonance luminous microspheres and application
CN102604305A (en) * 2012-02-21 2012-07-25 苏州纳诺康生物技术有限公司 Nano-grade fluorescent microspheres, preparation method thereof, and purpose thereof as markers used in immunity chromatography method
CN103333299A (en) * 2013-07-12 2013-10-02 西南大学 Glycidyl methacrylate and poly (glycol) methacrylate copolymer microsphere, preparation method and application of copolymer microsphere
CN106380617A (en) * 2016-09-06 2017-02-08 北京华科泰生物技术有限公司 Surface-activated fluorescent latex microspheres as well as preparation method and application thereof
CN106633168A (en) * 2016-10-19 2017-05-10 扬州倍赛德生物科技有限公司 Preparation method of monodisperse and large-particle-diameter polymer microspheres
CN112341663A (en) * 2020-10-28 2021-02-09 苏州纳微科技股份有限公司 ProteinA affinity chromatography medium of PMMA matrix and preparation method and application thereof

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
PRYMON, S等: ""Effect of Fe3O4 particles on multi-hollow morphology of poly(HEMA-divinylbenzene-styrene) microspheres prepared by Pickering suspension polymerization"", 《EXPRESS POLYMER LETTERS》 *
余丽丽等: ""盐酸小檗碱β-环糊精聚合物微球的制备和释药性能研究"", 《化工新型材料》 *
王晓明等: ""聚β-环糊精微球的制备及结构表征"", 《广东药学院学报》 *

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