CN113667048A - Monodisperse polymer color microsphere and preparation method and application thereof - Google Patents
Monodisperse polymer color microsphere and preparation method and application thereof Download PDFInfo
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- CN113667048A CN113667048A CN202110934220.1A CN202110934220A CN113667048A CN 113667048 A CN113667048 A CN 113667048A CN 202110934220 A CN202110934220 A CN 202110934220A CN 113667048 A CN113667048 A CN 113667048A
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- dye
- microsphere
- microspheres
- swelling
- carboxyl
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- 239000004005 microsphere Substances 0.000 title claims abstract description 158
- 229920000642 polymer Polymers 0.000 title claims abstract description 24
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- 238000000034 method Methods 0.000 claims abstract description 52
- 230000008961 swelling Effects 0.000 claims abstract description 49
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims abstract description 43
- 239000004793 Polystyrene Substances 0.000 claims abstract description 25
- 229920002223 polystyrene Polymers 0.000 claims abstract description 25
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 17
- 238000010556 emulsion polymerization method Methods 0.000 claims abstract description 5
- 239000000975 dye Substances 0.000 claims description 55
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 22
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 20
- 238000006243 chemical reaction Methods 0.000 claims description 19
- 239000000178 monomer Substances 0.000 claims description 18
- 238000006116 polymerization reaction Methods 0.000 claims description 17
- 238000003756 stirring Methods 0.000 claims description 17
- PPBRXRYQALVLMV-UHFFFAOYSA-N Styrene Chemical group C=CC1=CC=CC=C1 PPBRXRYQALVLMV-UHFFFAOYSA-N 0.000 claims description 16
- 239000006228 supernatant Substances 0.000 claims description 14
- 239000003999 initiator Substances 0.000 claims description 13
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 12
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 12
- 239000000839 emulsion Substances 0.000 claims description 11
- 239000003153 chemical reaction reagent Substances 0.000 claims description 10
- 239000008367 deionised water Substances 0.000 claims description 10
- 229910021641 deionized water Inorganic materials 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 9
- 238000010438 heat treatment Methods 0.000 claims description 8
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 6
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 6
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 claims description 6
- USHAGKDGDHPEEY-UHFFFAOYSA-L potassium persulfate Chemical compound [K+].[K+].[O-]S(=O)(=O)OOS([O-])(=O)=O USHAGKDGDHPEEY-UHFFFAOYSA-L 0.000 claims description 5
- NIXOWILDQLNWCW-UHFFFAOYSA-N 2-Propenoic acid Natural products OC(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 claims description 4
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 4
- ROOXNKNUYICQNP-UHFFFAOYSA-N ammonium persulfate Chemical compound [NH4+].[NH4+].[O-]S(=O)(=O)OOS([O-])(=O)=O ROOXNKNUYICQNP-UHFFFAOYSA-N 0.000 claims description 4
- 235000012730 carminic acid Nutrition 0.000 claims description 4
- 238000004587 chromatography analysis Methods 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 4
- TVMXDCGIABBOFY-UHFFFAOYSA-N octane Chemical compound CCCCCCCC TVMXDCGIABBOFY-UHFFFAOYSA-N 0.000 claims description 4
- SMZOUWXMTYCWNB-UHFFFAOYSA-N 2-(2-methoxy-5-methylphenyl)ethanamine Chemical compound COC1=CC=C(C)C=C1CCN SMZOUWXMTYCWNB-UHFFFAOYSA-N 0.000 claims description 3
- 239000006185 dispersion Substances 0.000 claims description 3
- -1 hexeneacetone Chemical compound 0.000 claims description 3
- 229910052757 nitrogen Inorganic materials 0.000 claims description 3
- 239000002244 precipitate Substances 0.000 claims description 3
- SCYULBFZEHDVBN-UHFFFAOYSA-N 1,1-Dichloroethane Chemical compound CC(Cl)Cl SCYULBFZEHDVBN-UHFFFAOYSA-N 0.000 claims description 2
- JKNCOURZONDCGV-UHFFFAOYSA-N 2-(dimethylamino)ethyl 2-methylprop-2-enoate Chemical compound CN(C)CCOC(=O)C(C)=C JKNCOURZONDCGV-UHFFFAOYSA-N 0.000 claims description 2
- XSHISXQEKIKSGC-UHFFFAOYSA-N 2-aminoethyl 2-methylprop-2-enoate;hydron;chloride Chemical compound Cl.CC(=C)C(=O)OCCN XSHISXQEKIKSGC-UHFFFAOYSA-N 0.000 claims description 2
- PCUPXNDEQDWEMM-UHFFFAOYSA-N 3-(diethylamino)propyl 2-methylprop-2-enoate Chemical compound CCN(CC)CCCOC(=O)C(C)=C PCUPXNDEQDWEMM-UHFFFAOYSA-N 0.000 claims description 2
- WWJCRUKUIQRCGP-UHFFFAOYSA-N 3-(dimethylamino)propyl 2-methylprop-2-enoate Chemical compound CN(C)CCCOC(=O)C(C)=C WWJCRUKUIQRCGP-UHFFFAOYSA-N 0.000 claims description 2
- UFQHFMGRRVQFNA-UHFFFAOYSA-N 3-(dimethylamino)propyl prop-2-enoate Chemical compound CN(C)CCCOC(=O)C=C UFQHFMGRRVQFNA-UHFFFAOYSA-N 0.000 claims description 2
- 235000009328 Amaranthus caudatus Nutrition 0.000 claims description 2
- 240000001592 Amaranthus caudatus Species 0.000 claims description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 2
- CERQOIWHTDAKMF-UHFFFAOYSA-N Methacrylic acid Chemical compound CC(=C)C(O)=O CERQOIWHTDAKMF-UHFFFAOYSA-N 0.000 claims description 2
- RGCKGOZRHPZPFP-UHFFFAOYSA-N alizarin Chemical compound C1=CC=C2C(=O)C3=C(O)C(O)=CC=C3C(=O)C2=C1 RGCKGOZRHPZPFP-UHFFFAOYSA-N 0.000 claims description 2
- 235000012735 amaranth Nutrition 0.000 claims description 2
- 239000004178 amaranth Substances 0.000 claims description 2
- 229910001870 ammonium persulfate Inorganic materials 0.000 claims description 2
- 239000000987 azo dye Substances 0.000 claims description 2
- ZFMQKOWCDKKBIF-UHFFFAOYSA-N bis(3,5-difluorophenyl)phosphane Chemical compound FC1=CC(F)=CC(PC=2C=C(F)C=C(F)C=2)=C1 ZFMQKOWCDKKBIF-UHFFFAOYSA-N 0.000 claims description 2
- 229950005499 carbon tetrachloride Drugs 0.000 claims description 2
- VYXSBFYARXAAKO-WTKGSRSZSA-N chembl402140 Chemical compound Cl.C1=2C=C(C)C(NCC)=CC=2OC2=C\C(=N/CC)C(C)=CC2=C1C1=CC=CC=C1C(=O)OCC VYXSBFYARXAAKO-WTKGSRSZSA-N 0.000 claims description 2
- 229960001701 chloroform Drugs 0.000 claims description 2
- 238000004140 cleaning Methods 0.000 claims description 2
- 150000001875 compounds Chemical class 0.000 claims description 2
- IQFVPQOLBLOTPF-HKXUKFGYSA-L congo red Chemical compound [Na+].[Na+].C1=CC=CC2=C(N)C(/N=N/C3=CC=C(C=C3)C3=CC=C(C=C3)/N=N/C3=C(C4=CC=CC=C4C(=C3)S([O-])(=O)=O)N)=CC(S([O-])(=O)=O)=C21 IQFVPQOLBLOTPF-HKXUKFGYSA-L 0.000 claims description 2
- 238000001816 cooling Methods 0.000 claims description 2
- LDHQCZJRKDOVOX-NSCUHMNNSA-N crotonic acid Chemical compound C\C=C\C(O)=O LDHQCZJRKDOVOX-NSCUHMNNSA-N 0.000 claims description 2
- 239000000463 material Substances 0.000 claims description 2
- 235000006408 oxalic acid Nutrition 0.000 claims description 2
- 239000012286 potassium permanganate Substances 0.000 claims description 2
- 239000001044 red dye Substances 0.000 claims description 2
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 claims description 2
- 238000000926 separation method Methods 0.000 claims description 2
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 claims description 2
- 235000019345 sodium thiosulphate Nutrition 0.000 claims description 2
- 239000002904 solvent Substances 0.000 claims description 2
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 claims description 2
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 2
- LDHQCZJRKDOVOX-UHFFFAOYSA-N trans-crotonic acid Natural products CC=CC(O)=O LDHQCZJRKDOVOX-UHFFFAOYSA-N 0.000 claims description 2
- 238000001514 detection method Methods 0.000 abstract description 30
- 238000012360 testing method Methods 0.000 abstract description 20
- 238000003317 immunochromatography Methods 0.000 abstract description 3
- 239000004816 latex Substances 0.000 abstract description 3
- 229920000126 latex Polymers 0.000 abstract description 3
- 238000007720 emulsion polymerization reaction Methods 0.000 abstract description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 16
- 238000005859 coupling reaction Methods 0.000 description 15
- 239000000243 solution Substances 0.000 description 15
- 230000008878 coupling Effects 0.000 description 14
- 238000010168 coupling process Methods 0.000 description 14
- 238000003908 quality control method Methods 0.000 description 12
- 239000000427 antigen Substances 0.000 description 11
- 102000036639 antigens Human genes 0.000 description 11
- 108091007433 antigens Proteins 0.000 description 11
- QYYMDNHUJFIDDQ-UHFFFAOYSA-N 5-chloro-2-methyl-1,2-thiazol-3-one;2-methyl-1,2-thiazol-3-one Chemical compound CN1SC=CC1=O.CN1SC(Cl)=CC1=O QYYMDNHUJFIDDQ-UHFFFAOYSA-N 0.000 description 8
- 229920001213 Polysorbate 20 Polymers 0.000 description 8
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 8
- 239000007983 Tris buffer Substances 0.000 description 8
- 239000007853 buffer solution Substances 0.000 description 8
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 8
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 8
- 230000008569 process Effects 0.000 description 8
- 239000011780 sodium chloride Substances 0.000 description 8
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 8
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 6
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 6
- 230000003213 activating effect Effects 0.000 description 6
- 230000004913 activation Effects 0.000 description 6
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 6
- 208000037797 influenza A Diseases 0.000 description 6
- 238000002156 mixing Methods 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- 125000000524 functional group Chemical group 0.000 description 5
- 239000012528 membrane Substances 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 238000005119 centrifugation Methods 0.000 description 4
- 238000000576 coating method Methods 0.000 description 4
- 239000003085 diluting agent Substances 0.000 description 4
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 4
- 229910000397 disodium phosphate Inorganic materials 0.000 description 4
- 235000019441 ethanol Nutrition 0.000 description 4
- 239000002245 particle Substances 0.000 description 4
- 239000003761 preservation solution Substances 0.000 description 4
- 238000001179 sorption measurement Methods 0.000 description 4
- 238000005303 weighing Methods 0.000 description 4
- 241001678559 COVID-19 virus Species 0.000 description 3
- 241000712431 Influenza A virus Species 0.000 description 3
- 241000701076 Macacine alphaherpesvirus 1 Species 0.000 description 3
- 230000004520 agglutination Effects 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
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- 241000283707 Capra Species 0.000 description 2
- 102000011632 Caseins Human genes 0.000 description 2
- 108010076119 Caseins Proteins 0.000 description 2
- 241000711573 Coronaviridae Species 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 229910021538 borax Inorganic materials 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- UQGFMSUEHSUPRD-UHFFFAOYSA-N disodium;3,7-dioxido-2,4,6,8,9-pentaoxa-1,3,5,7-tetraborabicyclo[3.3.1]nonane Chemical compound [Na+].[Na+].O1B([O-])OB2OB([O-])OB1O2 UQGFMSUEHSUPRD-UHFFFAOYSA-N 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 238000004043 dyeing Methods 0.000 description 2
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 2
- 108010046023 influenza B virus nucleoprotein Proteins 0.000 description 2
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- 108090000623 proteins and genes Proteins 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 229940080237 sodium caseinate Drugs 0.000 description 2
- 235000010339 sodium tetraborate Nutrition 0.000 description 2
- 239000004328 sodium tetraborate Substances 0.000 description 2
- RPENMORRBUTCPR-UHFFFAOYSA-M sodium;1-hydroxy-2,5-dioxopyrrolidine-3-sulfonate Chemical compound [Na+].ON1C(=O)CC(S([O-])(=O)=O)C1=O RPENMORRBUTCPR-UHFFFAOYSA-M 0.000 description 2
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- 101900222562 Influenza A virus Nucleoprotein Proteins 0.000 description 1
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- 239000002250 absorbent Substances 0.000 description 1
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- 239000000654 additive Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000013522 chelant Substances 0.000 description 1
- 238000012412 chemical coupling Methods 0.000 description 1
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- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000000986 disperse dye Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- OGPBJKLSAFTDLK-UHFFFAOYSA-N europium atom Chemical compound [Eu] OGPBJKLSAFTDLK-UHFFFAOYSA-N 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
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- 239000010931 gold Substances 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F212/00—Copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by an aromatic carbocyclic ring
- C08F212/02—Monomers containing only one unsaturated aliphatic radical
- C08F212/04—Monomers containing only one unsaturated aliphatic radical containing one ring
- C08F212/06—Hydrocarbons
- C08F212/08—Styrene
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
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- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Hematology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Food Science & Technology (AREA)
- Biotechnology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Cell Biology (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Processes Of Treating Macromolecular Substances (AREA)
Abstract
The invention discloses a monodisperse polymer color microsphere and a preparation method and application thereof, wherein the preparation method comprises the following steps: (1) preparing carboxyl polystyrene microspheres by adopting a soap-free emulsion polymerization method; (2) providing a swelling agent system having a dye dissolved therein; (3) and (3) permeating the dye into the interior of the carboxyl microsphere by a seed swelling method to obtain the colorful carboxyl polystyrene microsphere with the dye wrapped in the interior of the microsphere. The method adopts a one-step swelling method to obtain the functional microspheres embedded with the color dye, the seed microspheres used for swelling are carboxyl latex microspheres obtained in a soap-free emulsion polymerization mode, the operation is simple, the size of the obtained microspheres is uniform and controllable, and the surfaces of the microspheres are regular; can be used for immunochromatography detection test strip application.
Description
Technical Field
The invention relates to a monodisperse polymer color microsphere and a preparation method and application thereof.
Background
The polymer microsphere has the advantages of large surface area, good monodispersity, functionalized surface, strong adsorption and cohesion, stable morphological structure and the like, and is widely concerned in the fields of biomedicine, life science, chemical industry, microelectronic industry and the like. In the polymer microspheres, the preparation of polystyrene microspheres is different from the general polymerization process, and focuses on the final form of the polymer, and microspheres with different particle size distributions can be obtained by selecting different polymerization methods or by adjusting the reaction conditions. In recent years, the technology of the microsphere technology has developed rapidly, and the related latex agglutination reaction (LAT) is a specific agglutination generated by coupling an antibody (or antigen) on the microsphere surface and a corresponding antigen (or antibody), so that the detection is carried out by visual observation. However, since the content of antibody (or antigen) generated in the early stage is low, the contrast against the background is not obvious due to the white color of the obtained small amount of specific agglutination of the polymer microspheres, and the diagnosis is difficult to be determined by naked eyes, so that the application of the microspheres in the early stage of disease diagnosis such as inflammatory virus is limited. Therefore, the contrast is enhanced by dyeing the microspheres, so that the colored microspheres are easy to observe, thereby facilitating the improvement of the detection sensitivity.
The synthesis of the colored microspheres mainly lies in a combination mode of the microspheres and dye, and at present, the method is mainly divided into two preparation modes of a physical method and a chemical method, including a polymerization coating method, an embedding method, a direct adsorption method, a surface modification method, a chemical bonding method and the like. The dye molecules are coated at the outer end of the microsphere by a surface adsorption method, so that the loss of functional groups can be caused, and the detection sensitivity is reduced. The surface modification method is characterized in that dye molecules react with functional groups on the surfaces of microspheres to achieve a modification effect, although the obtained microspheres are bright in color, chemical modification occupies too many surface functional groups, the defects in a direct adsorption method exist, the surfaces of colored microspheres obtained through chemical bonding are easy to adhere, and the microspheres with good monodispersity are difficult to obtain. The physical embedding swelling method adopts monomer or swelling agent to swell the seed crosslinking microsphere, so that the volume of the seed crosslinking microsphere is increased, gaps are formed among crosslinking molecules, disperse dye can be dissolved by taking the swelling agent as a medium, and the monomer carries the dye to continuously enter the gaps in the microsphere after the microsphere is swelled to be enlarged until the equilibrium state is finally reached. The dye is embedded in the polymer microsphere and is not easy to leak. The morphological structure of the microspheres is stable in the seed formation process, and when the swelling condition is properly controlled, the shape of the microspheres is not easy to change.
Most of the color microspheres obtained at present have color appearance and sphericity, and the surface functionalized particle size can be adjusted and compatible. The dye substance is coated inside the microsphere in a swelling mode, so that the structural form of the microsphere is maintained, the coating rate is adjustable, and the original form and the surface of the microsphere are not affected.
Disclosure of Invention
The invention aims to provide monodisperse polymer colored microspheres which are uniform in particle size, adjustable in surface carboxyl density, bright and full in appearance color and luster, and a preparation method and application thereof.
The technical solution of the invention is as follows:
a monodisperse polymer color microsphere is characterized in that: prepared by the following method; the preparation method comprises the following steps:
(1) preparing carboxyl polystyrene microspheres by adopting a soap-free emulsion polymerization method;
(2) providing a swelling agent system having a dye dissolved therein;
(3) and (3) permeating the dye into the interior of the carboxyl microsphere by a seed swelling method to obtain the colorful carboxyl polystyrene microsphere with the dye wrapped in the interior of the microsphere.
A preparation method of monodisperse polymer colored microspheres is characterized by comprising the following steps: comprises the following steps:
(1) preparing carboxyl polystyrene microspheres by adopting a soap-free emulsion polymerization method;
(2) providing a swelling agent system having a dye dissolved therein;
(3) and (3) permeating the dye into the interior of the carboxyl microsphere by a seed swelling method to obtain the colorful carboxyl polystyrene microsphere with the dye wrapped in the interior of the microsphere.
The step (1) comprises the following steps:
(a) taking a dispersion water system in a water bath, adding a styrene monomer and a functional monomer in sequence under the protection of nitrogen, and stirring uniformly;
(b) adding an initiator into a monomer system for dissolving;
(c) and after the materials are fully dissolved, opening a water bath for heating, and keeping the polymerization reaction after reaching the target temperature to obtain the carboxyl polystyrene microsphere seed emulsion.
The functional monomer is selected from one or more of the following compounds: methacrylic acid, 2-aminoethyl methacrylate hydrochloride, dimethylaminopropyl methacrylate, dimethylaminoethyl methacrylate, diethylaminopropyl methacrylate, dimethylaminopropyl acrylate, acrylic acid, crotonic acid;
the initiator is a water-phase initiator and is one or more of potassium persulfate, a mixture of potassium persulfate and sodium thiosulfate, and a mixture of ammonium persulfate or potassium permanganate and oxalic acid.
The amount of the initiator is 0.1-0.55 wt% of the amount of the styrene monomer; step (c), keeping the polymerization reaction for 4-24 h;
the polymerization reaction is carried out at a stirring speed of 200-600 rpm, and the polymerization temperature is 60-80 ℃; the interval time between the addition of the initiator and the start of heating is 30-60 min; the volume ratio of the styrene to the functional monomer is 0.1-0.4: 1.
the specific method of step (1) may be: firstly, adding deionized water with a certain volume into a 500 mL three-neck flask, introducing nitrogen, removing oxygen in a reaction system, adding styrene, then adding 1.1-4.4 mL of carboxyl functional monomer, uniformly stirring by an electronic digital display stirrer at a certain stirring speed, adding 0.1-0.55 wt% of initiator based on the amount of the styrene monomer into the monomer system after 15 min for dissolving, uniformly stirring, starting heating after 30 min, raising the temperature to the polymerization temperature, and keeping for 4-24 h to obtain the milky carboxyl polystyrene microsphere seed.
The swelling system dissolved with the dye is to dissolve the dye in a swelling agent under the ultrasonic condition; the dye is selected from one or a mixed type of water-soluble dye or oil-soluble dye;
the water-soluble dye may be selected from: active red dye, amaranth, carmine, rhodamine 6G, rhodamine and alizarin red;
the oil-soluble dye may be selected from: solvent red series, azo carmine, lipophilic azo dyes, congo red;
the swelling agent is selected from: tetrahydrofuran, tetrachloromethane, trichloromethane, dichloromethane, hexeneacetone, toluene, ethanol, dichloroethane, dioctyl adipate, n-octane, n-hexane or different combinations of the above.
The method in the step (3) comprises the following steps:
(a) adding the seed microspheres, a swelling reagent, a dye and deionized water into a three-neck flask, and heating to a swelling temperature under a constant stirring state;
(b) after the temperature is raised to the swelling temperature, cooling the system temperature to room temperature, carrying out solid-liquid separation in a centrifugal mode, and removing supernatant;
(c) and continuously adding a swelling reagent to disperse and dissolve redundant dyes, continuously centrifuging to remove supernatant, repeating the step for 2-3 times by using deionized water, and finally adding a small amount of deionized water to disperse and store the microspheres.
The swelling temperature is 50-75 ℃; the stirring speed of the swelling reaction is 200-500 rpm/min; the swelling reaction stirring time is 3-12 h; the volume of the carboxyl polystyrene microsphere seed emulsion is 0.2-1 time of that of the swelling medium; after the reaction is finished, removing supernatant through high-speed centrifugation to obtain microsphere precipitate, cleaning redundant dye by adopting a mild swelling agent to obtain colored microspheres, and performing dispersion storage by adopting deionized water.
The mild swelling agent is preferably a dissolved dye selected from: one or more of toluene, ethanol, ethyl acetate, acetone and dichloromethane.
The use of monodisperse polymer colored microspheres is characterized in that: as microsphere markers in immuno-lateral chromatography.
The invention adopts a one-step swelling method to obtain the functionalized polystyrene microspheres for embedding the target organic dye, the seed microspheres for swelling are carboxyl latex microspheres obtained in a soap-free emulsion polymerization mode, the operation is simple, the size of the obtained microspheres is uniform and controllable, and the surface is regular.
The beneficial effect of this patent still lies in:
1) the invention selects a polymerization mode of soap-free emulsion to obtain microsphere seeds with clean surfaces, avoids unnecessary variables generated in the subsequent swelling reaction of the seeds due to addition of excessive additives such as surfactants, stabilizers, hydrophobing agents and the like, and is difficult to clean. The preparation of the functional microspheres adopts a one-step polymerization method to introduce carboxyl groups on the surfaces of the microspheres, and the carboxyl density on the surfaces of the microspheres is adjusted by changing the using amount of functional monomers, and the shapes and monodispersity of the microspheres are not influenced; the size and monodispersity of the microspheres are regulated and controlled by regulating factors such as stirring speed, stirring temperature and the like.
2) The invention adopts a one-step swelling embedding mode to coat the target dye in the microsphere, avoids the defects generated by preparing the fluorescent microsphere by a physical and chemical method, ensures the stability of the dye, has no leakage defect in later application, and thus has influence on the application performance.
3) The polymer color functional microspheres prepared by the invention have the advantages of clean surface, uniform size distribution and bright appearance color.
4) The color functional microsphere prepared by the invention can be combined with an antibody (or antigen) through a surface active functional group to achieve the purpose of accurate quantification of a detection result by color tracing. The detection performance index of the method is far higher than that of the traditional rapid detection technology such as colloidal gold and the like, and the method has a great application prospect in the biomedical detection industry.
5) The color microsphere immunochromatography detection test strip is developed by adopting a double-antibody sandwich method mode, and the antibody is labeled by a chemical coupling method to form stable combination of the antibody and the microsphere, thereby realizing the advantage of high-sensitivity quantitative detection.
Drawings
The invention is further illustrated by the following figures and examples.
FIG. 1 is an infrared spectrum of surface functionalized polystyrene microspheres.
FIG. 2 is an infrared spectrum of surface functionalized colored polystyrene microspheres.
FIG. 3 is a scanning electron microscope image of polystyrene seed microspheres prepared from soap-free emulsion.
FIG. 4 is a scanning electron microscope image of the functionalized polystyrene microspheres after dyeing.
FIG. 5 is a 2019-nCoV novel coronavirus antigen chromatographic test strip.
In FIG. 6, the test strip chromatography results of the color microspheres obtained by coupling antibody show that the antigen quality control concentrations from left to right are 50 pg/mL, 100 pg/mL, 250 pg/mL and 500 pg/mL, respectively.
FIG. 7 is an influenza A/B virus antigen immunochromatographic test strip.
FIG. 8 shows the test strip of the colored microsphere coupled with the antibody, the commercial microsphere is on the left, the self-made microsphere is on the right, and the concentrations of the antigen reference products from left to right are 1 (1: 500), 1 (1: 1000), 1 (1: 2000) and 1 (1: 4000), respectively.
Detailed Description
Reagent and apparatus
Styrene (Styrene, St), Acrylic acid (Acrylic, AA), absolute ethyl alcohol (Ethanol), Potassium persulfate (KPS) and dyes are analytically pure, and are purchased from national drug group chemical reagent company Limited, and water used in laboratories is deionized water.
The invention provides a method for preparing a colored polystyrene microsphere and application thereof, and particularly relates to the colored polystyrene microsphere prepared by the method, which is bright in color, uniform in appearance and appearance, good in monodispersity, good in stability and simple in preparation process.
The following detailed description of embodiments of the invention refers to the accompanying drawings. Specific examples the technical solution of the present invention is implemented on the premise of detailed implementation and specific operation, but the scope of the present invention is not limited to the following examples.
Example 1
Synthesizing functional polystyrene seed microspheres by adopting a soap-free emulsion synthesis mode; controlling the size of the microspheres to be 100-400 nm by adjusting parameters of a reaction system, such as the dosage of a polymerization monomer, a functional monomer and an initiator, polymerization temperature, stirring speed and the like; the density of carboxyl on the surface of the microsphere is 60-300 mu mol/g;
the method specifically comprises the following steps: mechanically stirring a three-neck flask filled with a polymerization monomer and a functional monomer in a water bath heating tank at a certain temperature (100-500 rpm/min), adding 0.1-0.55 wt% of an initiator dissolved in water in advance based on the dosage of a styrene monomer, wherein no excess emulsifier is added, and mixing water or water and ethanol according to a certain volume ratio except for the reaction monomer in a reaction combination manner.
Weighing 60 mg of target dye A and 60 mg of target dye B, dissolving the dyes A and B in a three-neck flask filled with 50mL of absolute ethyl alcohol, adopting an ultrasonic dissolving mode to fully dissolve the dyes in a reaction medium, directly weighing more than 5 mL of the obtained microsphere seed white emulsion without further treatment, ultrasonically dispersing the white emulsion in the three-neck flask, uniformly mixing the seed microspheres, the reaction medium and the dyes by using a rotating speed of 100-500 rpm/min, and reacting for at least 5 hours to obtain the colored microspheres with the appearances shown in figure 8.
Example 2
Weighing 50 mg of target dye A and 50 mg of target dye B, dissolving the target dye A and the target dye B in a three-neck flask filled with 100mL of absolute ethyl alcohol, adopting an ultrasonic dissolving mode to fully dissolve the dyes in the absolute ethyl alcohol, directly weighing more than 5 mL of the obtained microsphere seed white emulsion without further treatment, ultrasonically dispersing the microsphere seed white emulsion in the three-neck flask, uniformly mixing the seed microspheres, a reaction medium and the dyes by using a rotating speed of 200-500 rpm/min, and reacting for at least 7 hours.
The diameter of the microsphere seed obtained in this embodiment is 300 nm, as shown in FIG. 3, and the nano size after swelling does not change significantly, as shown in FIG. 2.
The surface functional groups of the microspheres in this embodiment are unchanged before and after swelling, as can be seen from the infrared spectrum in fig. 1.
The application case one:
the application case provides a carboxyl color microsphere coupling antibody process and a 2019-nCoV novel coronavirus antigen immunochromatography detection test strip:
the test strip of the invention is composed of four parts as shown in figure 5: sample pad, conjugate pad, NC membrane and absorbent pad. The test paper is stuck on a PVC bottom plate in a mutual overlapping mode (the overlapping part is about 2 mm) according to a certain sequence, and a complete test paper strip system is formed after the test paper strip is assembled. As shown in FIG. 4, a membrane drawing gold spraying instrument is used for drawing lines on an NC membrane, and monoclonal N antibodies and goat anti-mouse IgG antibodies are coated as detection lines (T lines) and quality control lines (C lines) of the test strip. Because the particle size of the color nanoprobe used by the test strip detection and the property of a detected object directly influence the chromatography rate and the fluency of the color nanoprobe on the test strip, in order to reduce batch-to-batch difference in the test strip detection process, the test strip can carry out corresponding optimization treatment such as optimization of a binding pad, activation of a glass fiber membrane and the like according to specific experimental conditions.
The preparation of the main solution required by the colored microsphere marker sprayed on the bonding pad in the process of coupling and marking the antibody is as follows:
1) carboxyl color nano microsphere activation buffer solution: 50 mM MES, pH 6.0;
2) carboxyl color nano microsphere activating reagent a: MES solution as above containing 1% EDC;
3) and (b) a carboxyl color nano microsphere activating reagent: MES solution containing 1% of sulfo-NHS;
4) coupling the carboxyl color nano microspheres with protein buffer solution: 25 mM PB, pH 7.0;
5) washing liquid: 25 mM Tris-base, 0.2% Tween 20, 0.15M NaCl, 0.05% ProClin 300, pH 7.8;
6) sealing liquid: 50 mM PB, 5% BSA, pH 8.0;
7) preservation solution: 25 mM Tris-base, 0.05% Tween 20, 0.15M NaCl, 0.05% ProClin 300, 1% BSA, 5% trehalose, pH 7.2;
8) marking microsphere diluent: 25 mM Tris-base, 0.05% Tween 20, 0.05% ProClin 300, 1% BSA, 5% trehalose, 20% sucrose, pH 9.0;
9) coating liquid: 10 mM Na2HPO4·12H2O, 0.15M NaCl, 0.3% trehalose, 0.1% NaN3，pH7.4；
10) Sample pad pretreatment solution: 10 mM sodium tetraborate, 1% PVP, 0.2% sodium caseinate, 1% Trition-X100, 1% S9, 0.02% NaN3；
11) Binding pad pretreatment solution: 50 mM Na2HPO4·12H2O，1% Trition-X100，0.5% PVA，0.5% BSA，pH 7.4；
12) And (3) calibrating product diluent: 50 mM Tris-base, 0.15M NaCl, 0.1% ProClin 300, 0.01% Tween-20, 1.5% BSA, pH 7.8;
the method for coupling and marking the antibody by the colored functional microspheres comprises the following specific implementation steps:
1) pretreating carboxylated color nano microspheres:
taking 50 mu L (2 mg) of a solution with 4% solid content, centrifuging the solution in a 1mL centrifuge tube at 4 ℃ for 15 min at the centrifugation speed of 15000 rpm/min, and carefully leaving supernatant; the process is repeated for 2 times to ensure that impurities on the surface of the colored microspheres are removed.
2) Activation of the carboxylated color nano microspheres:
a) adding 1mL of carboxylated color nano microsphere activation buffer solution into the centrifugal precipitate, fully and uniformly suspending the fluorescent microspheres by using a vortex mixer, centrifuging at the centrifugal speed of 15000 rpm/min for 15 min at 4 ℃, and carefully removing supernatant; repeating the process for 1 time, adding 1mL of carboxylated color nano microsphere activation buffer solution, and uniformly mixing and resuspending again;
b) adding 4.8 μ L of activating agent a and 48 μ L of activating agent b, ultrasonic resuspending, uniformly dispersing with cell ultrasonic disruptor under ice bath at power of 5%, working for 1s, spacing 3s, and lasting for 1 min, and rotating on vertical mixer at room temperature for 30 min. Centrifuging at the temperature of 4 ℃ at the centrifugal speed of 15000 rpm/min for 15 min, and carefully leaving the supernatant;
c) adding 1ml of microsphere coupling buffer solution, fully and uniformly suspending the color microspheres by using a vortex mixer, ultrasonically resuspending (under the conditions as above), centrifuging at the centrifugal rate of 15000 rpm/min for 15 min at 4 ℃, and carefully removing supernatant; this procedure was repeated 1 time.
3) Coupling the carboxylated color nano microspheres with an antibody:
a) and adding 1mL of microsphere coupling buffer solution, then carrying out ultrasonic resuspension, adding 50 mu L (2.4 mg/mL) of N28 new crown antibody, fully and uniformly mixing the suspended carboxyl color nano microspheres by using a vortex mixer, and carrying out coupling reaction for 2 hours at the rotation condition of 37 ℃. Centrifuging at 4 ℃ at a centrifugation speed of 10000 rpm/min for 15 min, and carefully removing the supernatant;
b) adding 1mL of microsphere washing solution, carrying out ultrasonic resuspension, centrifuging at 4 ℃ at a centrifugation speed of 10000 rpm/min for 15 min, and carefully removing a supernatant; this procedure was repeated 1 time.
4) And (3) sealing and storing the carboxylated color nano microspheres:
a) adding 1mL of confining liquid, carrying out ultrasonic resuspension on the carboxyl color nano microspheres, fully and uniformly mixing the suspended carboxyl color nano microspheres by using a vortex mixer, and rotating and confining for 1 hour.
b) Centrifuging the microsphere suspension of the centrifugal tube with the volume of 1.5 mL for 15 min at the centrifugal rate of 10000 rpm/min at the temperature of 4 ℃, and carefully removing supernatant;
c) adding 1mL of preservation solution, ultrasonically resuspending the carboxyl color nano microspheres, centrifuging at 4 ℃ at a centrifugation rate of 10000 rpm/min for 15 min, and carefully removing supernatant; repeating the step for 1 time;
d) diluting the marked carboxylated europium chelate colored nano-microspheres to 2 mg/mL by using a preservation solution, storing at 2-8 ℃, and keeping in the dark for later use.
The invention has the following analysis for the detection result:
when the detection result is positive, the displayed strip line image shows two color reaction lines, one is in the T area (detection line) and the other is in the C area (quality control line)
When the detection result is positive, the presented strip line image shows that only one bright reaction line appears in the quality control line C area, and no visible red strip exists in the detection area.
And (4) analyzing results:
as can be seen from FIG. 6The color polystyrene carboxyl microsphere prepared by the invention is coupled with a new crown N28 antibody through a series of optimized condition control experiments, is used for detecting novel coronavirus SARS-CoV-2, has the quality control product concentrations of 50 pg/mL, 100 pg/mL, 250 pg/mL and 500 pg/mL respectively from left to right, and has the signal-to-noise ratio (S) along with the increase of the quality control product concentrationT/SC) Gradually increasing. The results show that the test strip has high sensitivity and high specificity.
Application case two:
the application case provides a carboxyl color microsphere coupling antibody process and an influenza A/B virus antigen detection test strip:
the structure of the test strip is consistent with that of the first embodiment, and the specific structure is shown in figure 7;
an influenza A/B virus antigen detection test strip comprises detection lines A and B and a quality control line C on an NC membrane, wherein the detection line A is coated with influenza A and/or influenza B virus NP protein monoclonal antibodies, the detection line B is coated with influenza B virus NP protein monoclonal antibodies, and the quality control line C is coated with goat anti-mouse IgG.
The preparation of the main solution required by the color functional microsphere marker sprayed on the bonding pad in the process of coupling and marking the antibody is as follows:
1) carboxyl color nano microsphere activation buffer solution: 50 mM MES, pH 6.0;
2) carboxyl color nano microsphere activating reagent a: MES solution as above containing 1% EDC;
3) and (b) a carboxyl color nano microsphere activating reagent: MES solution containing 1% of sulfo-NHS;
4) coupling the carboxyl color nano microspheres with protein buffer solution: 25 mM PB, pH 7.0;
5) washing liquid: 25 mM Tris-base, 0.02% Tween 20, 0.15M NaCl, 0.05% ProClin 300, pH 7.8;
6) sealing liquid: 50 mM PB, 5% BSA, pH 8.0;
7) preservation solution: 25 mM Tris-base, 0.05% Tween 20, 0.15M NaCl, 0.05% ProClin 300, 1% BSA, 5% trehalose, pH 7.2;
8) marking microsphere diluent: 25 mM Tris-base, 0.05% Tween 20, 0.05% ProClin 300, 1% BSA, 5% trehalose, 20% sucrose, Ph9.0;
9) coating liquid: 10 mM Na2HPO4·12H2O, 0.15M NaCl, 0.3% trehalose, 0.1% NaN3，pH7.4；
10) Sample pad pretreatment solution: 10 mM sodium tetraborate, 1% PVP, 0.2% sodium caseinate, 1% Trition-X100, 1% S9, 0.02% NaN3；
11) Binding pad pretreatment solution: 50 mM Na2HPO4·12H2O，1% Trition-X100，0.5% PVA，0.5% BSA，pH 7.4；
12) And (3) calibrating product diluent: 50 mM Tris-base, 0.15M NaCl, 0.1% ProClin 300, 0.01% Tween-20, 1.5% BSA, pH 7.8;
the scheme and the specific implementation steps of the method for coupling and labeling the anti-fluA-NP-1 by the colored carboxyl microspheres are consistent with the scheme of coupling the N28 antibody in the implementation case one.
The detection results of the samples present an explanation:
positive for influenza A: two red reaction lines appeared, one in zone A (detection zone) and one in zone C (quality control zone).
Positive flow B: two red reaction lines appeared, one in zone B (detection zone) and one in zone C (quality control zone).
Positive for influenza A and B: three red reaction lines appeared, one in zone a (detection zone), one in zone B (detection zone), and one in zone C (quality control zone).
Negative: only one red reaction line appears in the quality control zone C, and no visible red strip appears in the detection zone.
Claims (10)
1. A monodisperse polymer color microsphere is characterized in that: prepared by the following method; the preparation method comprises the following steps:
(1) preparing carboxyl polystyrene microspheres by adopting a soap-free emulsion polymerization method;
(2) providing a swelling agent system having a dye dissolved therein;
(3) and (3) permeating the dye into the interior of the carboxyl microsphere by a seed swelling method to obtain the colorful carboxyl polystyrene microsphere with the dye wrapped in the interior of the microsphere.
2. A method for preparing monodisperse polymer colored microspheres according to claim 1, wherein the method comprises the following steps: comprises the following steps:
(1) preparing carboxyl polystyrene microspheres by adopting a soap-free emulsion polymerization method;
(2) providing a swelling agent system having a dye dissolved therein;
(3) and (3) permeating the dye into the interior of the carboxyl microsphere by a seed swelling method to obtain the colorful carboxyl polystyrene microsphere with the dye wrapped in the interior of the microsphere.
3. The method for preparing monodisperse polymer color microspheres according to claim 2, wherein: the step (1) comprises the following steps:
(a) taking a dispersion water system in a water bath, adding a styrene monomer and a functional monomer in sequence under the protection of nitrogen, and stirring uniformly;
(b) adding an initiator into a monomer system for dissolving;
(c) and after the materials are fully dissolved, opening a water bath for heating, and keeping the polymerization reaction after reaching the target temperature to obtain the carboxyl polystyrene microsphere seed emulsion.
4. The method for preparing monodisperse polymer color microspheres according to claim 3, wherein: the functional monomer is selected from one or more of the following compounds: methacrylic acid, 2-aminoethyl methacrylate hydrochloride, dimethylaminopropyl methacrylate, dimethylaminoethyl methacrylate, diethylaminopropyl methacrylate, dimethylaminopropyl acrylate, acrylic acid, crotonic acid;
the initiator is a water-phase initiator and is one or more of potassium persulfate, a mixture of potassium persulfate and sodium thiosulfate, and a mixture of ammonium persulfate or potassium permanganate and oxalic acid.
5. The method for preparing monodisperse polymer color microspheres according to claim 3, wherein: the amount of the initiator is 0.1-0.55 wt% of the amount of the styrene monomer; step (c), keeping the polymerization reaction for 4-24 h;
the polymerization reaction is carried out at a stirring speed of 200-600 rpm, and the polymerization temperature is 60-80 ℃; the interval time between the addition of the initiator and the start of heating is 30-60 min; the volume ratio of the styrene to the functional monomer is 0.1-0.4: 1.
6. the method for preparing monodisperse polymer color microspheres according to claim 2, wherein: the swelling system dissolved with the dye is to dissolve the dye in a swelling agent under the ultrasonic condition; the dye is selected from one or a mixed type of water-soluble dye or oil-soluble dye;
the water-soluble dye may be selected from: active red dye, amaranth, carmine, rhodamine 6G, rhodamine and alizarin red;
the oil-soluble dye may be selected from: solvent red series, azo carmine, lipophilic azo dyes, congo red;
the swelling agent is selected from: tetrahydrofuran, tetrachloromethane, trichloromethane, dichloromethane, hexeneacetone, toluene, ethanol, dichloroethane, dioctyl adipate, n-octane, n-hexane or different combinations of the above.
7. The method for preparing monodisperse polymer color microspheres according to claim 2, wherein: the method in the step (3) comprises the following steps:
(a) adding the seed microspheres, a swelling reagent, a dye and deionized water into a three-neck flask, and heating to a swelling temperature under a constant stirring state;
(b) after the temperature is raised to the swelling temperature, cooling the system temperature to room temperature, carrying out solid-liquid separation in a centrifugal mode, and removing supernatant;
(c) and continuously adding a swelling reagent to disperse and dissolve redundant dyes, continuously centrifuging to remove supernatant, repeating the step for 2-3 times by using deionized water, and finally adding a small amount of deionized water to disperse and store the microspheres.
8. The method for preparing monodisperse polymer color microspheres of claim 7, wherein: the swelling temperature is 50-75 ℃; the stirring speed of the swelling reaction is 200-500 rpm/min; the swelling reaction stirring time is 3-12 h; the volume of the carboxyl polystyrene microsphere seed emulsion is 0.2-1 time of that of the swelling medium; after the reaction is finished, centrifuging to remove supernatant to obtain microsphere precipitate, cleaning redundant dye by adopting a mild swelling agent to obtain colored microspheres, and dispersing and storing by adopting deionized water.
9. The method for preparing monodisperse polymer color microspheres of claim 8, wherein: the mild swelling agent is preferably a dissolved dye selected from: one or more of toluene, ethanol, ethyl acetate, acetone and dichloromethane.
10. Use of monodisperse polymer colour microspheres according to claim 1, characterised in that: as microsphere markers in immuno-lateral chromatography.
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| CN114933897A (en) * | 2022-03-11 | 2022-08-23 | 江西维邦生物科技有限公司 | Method for preparing aggregation-induced emission microspheres by swelling method and application thereof |
| CN116550311A (en) * | 2023-07-10 | 2023-08-08 | 南京立顶医疗科技有限公司 | Method for synthesizing color microspheres by hydrothermal synthesis method and application |
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