CN112098639B - Synthesis and application of secondary antibody with graphene oxide as carrier - Google Patents
Synthesis and application of secondary antibody with graphene oxide as carrier Download PDFInfo
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- CN112098639B CN112098639B CN202010991925.2A CN202010991925A CN112098639B CN 112098639 B CN112098639 B CN 112098639B CN 202010991925 A CN202010991925 A CN 202010991925A CN 112098639 B CN112098639 B CN 112098639B
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- graphene oxide
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- phenolphthalein
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 title claims abstract description 34
- 229910021389 graphene Inorganic materials 0.000 title claims abstract description 34
- 230000015572 biosynthetic process Effects 0.000 title abstract description 7
- 238000003786 synthesis reaction Methods 0.000 title abstract description 7
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 14
- 238000000034 method Methods 0.000 claims abstract description 11
- 238000002965 ELISA Methods 0.000 claims abstract description 10
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000004327 boric acid Substances 0.000 claims abstract description 8
- 238000011068 loading method Methods 0.000 claims abstract description 4
- KJFMBFZCATUALV-UHFFFAOYSA-N phenolphthalein Chemical compound C1=CC(O)=CC=C1C1(C=2C=CC(O)=CC=2)C2=CC=CC=C2C(=O)O1 KJFMBFZCATUALV-UHFFFAOYSA-N 0.000 claims description 20
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 17
- 239000000243 solution Substances 0.000 claims description 13
- NAQMVNRVTILPCV-UHFFFAOYSA-N hexane-1,6-diamine Chemical compound NCCCCCCN NAQMVNRVTILPCV-UHFFFAOYSA-N 0.000 claims description 12
- SXGZJKUKBWWHRA-UHFFFAOYSA-N 2-(N-morpholiniumyl)ethanesulfonate Chemical compound [O-]S(=O)(=O)CC[NH+]1CCOCC1 SXGZJKUKBWWHRA-UHFFFAOYSA-N 0.000 claims description 10
- -1 polyoxyethylene Polymers 0.000 claims description 10
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 9
- LDKDGDIWEUUXSH-UHFFFAOYSA-N Thymophthalein Chemical compound C1=C(O)C(C(C)C)=CC(C2(C3=CC=CC=C3C(=O)O2)C=2C(=CC(O)=C(C(C)C)C=2)C)=C1C LDKDGDIWEUUXSH-UHFFFAOYSA-N 0.000 claims description 9
- 229940098773 bovine serum albumin Drugs 0.000 claims description 9
- 239000008055 phosphate buffer solution Substances 0.000 claims description 9
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 claims description 8
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 claims description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 7
- 229920003171 Poly (ethylene oxide) Polymers 0.000 claims description 6
- 150000004985 diamines Chemical class 0.000 claims description 6
- NQBHBNIZBDDDRH-UHFFFAOYSA-N (2,6-difluoro-4-formylphenyl)boronic acid Chemical compound OB(O)C1=C(F)C=C(C=O)C=C1F NQBHBNIZBDDDRH-UHFFFAOYSA-N 0.000 claims description 5
- 238000006243 chemical reaction Methods 0.000 claims description 5
- 239000003153 chemical reaction reagent Substances 0.000 claims description 5
- BEOOHQFXGBMRKU-UHFFFAOYSA-N sodium cyanoborohydride Chemical compound [Na+].[B-]C#N BEOOHQFXGBMRKU-UHFFFAOYSA-N 0.000 claims description 5
- PVAWONNALJEQJH-UHFFFAOYSA-N (2,4-difluoro-3-formylphenyl)boronic acid Chemical compound OB(O)C1=CC=C(F)C(C=O)=C1F PVAWONNALJEQJH-UHFFFAOYSA-N 0.000 claims description 4
- 239000004698 Polyethylene Substances 0.000 claims description 4
- 239000006185 dispersion Substances 0.000 claims description 4
- SWRGUMCEJHQWEE-UHFFFAOYSA-N ethanedihydrazide Chemical compound NNC(=O)C(=O)NN SWRGUMCEJHQWEE-UHFFFAOYSA-N 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 229920000573 polyethylene Polymers 0.000 claims description 4
- 239000002994 raw material Substances 0.000 claims description 4
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 claims description 3
- LIUCWHQVLKSECA-UHFFFAOYSA-N 2-hydroxyacetohydrazide Chemical compound NNC(=O)CO LIUCWHQVLKSECA-UHFFFAOYSA-N 0.000 claims description 2
- FCWAOWVIYGJOPZ-UHFFFAOYSA-N 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-2-carbaldehyde Chemical compound O1C(C)(C)C(C)(C)OB1C1=CC=C(C=O)N=C1 FCWAOWVIYGJOPZ-UHFFFAOYSA-N 0.000 claims description 2
- 230000004913 activation Effects 0.000 claims description 2
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 2
- 230000002209 hydrophobic effect Effects 0.000 claims description 2
- 229910000403 monosodium phosphate Inorganic materials 0.000 claims description 2
- 235000019799 monosodium phosphate Nutrition 0.000 claims description 2
- 238000002414 normal-phase solid-phase extraction Methods 0.000 claims description 2
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 2
- 238000007142 ring opening reaction Methods 0.000 claims description 2
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 claims description 2
- 238000010189 synthetic method Methods 0.000 claims description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 2
- 239000007853 buffer solution Substances 0.000 claims 1
- ZOXJGFHDIHLPTG-UHFFFAOYSA-N Boron Chemical compound [B] ZOXJGFHDIHLPTG-UHFFFAOYSA-N 0.000 abstract description 4
- 229910052796 boron Inorganic materials 0.000 abstract description 4
- 125000006850 spacer group Chemical group 0.000 abstract description 4
- 238000007306 functionalization reaction Methods 0.000 abstract description 3
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 238000004321 preservation Methods 0.000 abstract description 2
- 150000001720 carbohydrates Chemical class 0.000 abstract 1
- HEMHJVSKTPXQMS-UHFFFAOYSA-M sodium hydroxide Inorganic materials [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 8
- 238000005119 centrifugation Methods 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 102000004338 Transferrin Human genes 0.000 description 4
- 108090000901 Transferrin Proteins 0.000 description 4
- 239000012581 transferrin Substances 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- 239000000872 buffer Substances 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 238000001308 synthesis method Methods 0.000 description 2
- 238000008157 ELISA kit Methods 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 239000007987 MES buffer Substances 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 239000012670 alkaline solution Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003124 biologic agent Substances 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 230000031018 biological processes and functions Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- PWYPGEFOZYBWDP-UHFFFAOYSA-N boric acid;pyridine Chemical class OB(O)O.C1=CC=NC=C1 PWYPGEFOZYBWDP-UHFFFAOYSA-N 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 150000002009 diols Chemical class 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000009533 lab test Methods 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
Classifications
-
- 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
- G01N33/531—Production of immunochemical test materials
Abstract
The invention relates to synthesis and application of secondary antibody (secondary antibody) with graphene oxide as a carrier. The synthesis steps comprise: grafting a spacer on graphene oxide; boric acid functionalization is realized on graphene oxide; loading a pH sensitive color developing agent on the functional graphene oxide; covalent bonding is performed by utilizing boron affinity, and pH is adjusted to enable the color developing agent to develop color. The present invention is capable of binding to the cis-diol of the saccharide on an antibody under physiological pH conditions. The artificial secondary antibody solves the problems of high production cost, difficult preservation and transportation and poor stability of biological secondary antibodies, and is suitable for a common ELISA process.
Description
Technical Field
The invention relates to the field of biological materials, in particular to synthesis and application of secondary antibody (secondary antibody) taking graphene oxide as a carrier.
Background
Proteins are an important substance in biological processes of organisms, and in recent years, with the development of proteomics, proteins have been further known and understood. Protein is used as the basis of living matters and has close relation with the generation, development, diagnosis and treatment of diseases, so quantitative detection of the protein is an indispensable technical means, but the content of the protein in an actual sample is extremely low, and complex matrix interference exists, so that the quantitative determination of the protein is difficult and heavy. The development of enzyme-linked immunosorbent assay (ELISA) technology basically solves the problem, but the biological reagents used in the ELISA process are generally stored at low temperature for no more than 6 months, and the production is complex and the cost is high. Therefore, the search for an effective biological agent alternative is critical to effectively solve this problem. The preparation of the secondary antibody by adopting an artificial synthesis method is not reported at present.
Disclosure of Invention
The invention aims to provide synthesis and application of a secondary antibody with graphene oxide as a carrier, namely, a secondary antibody (artificial secondary antibody) is prepared by adopting an artificial synthesis method. By grafting a spacer onto graphene oxide and then boric acid functionalization, it is enabled to bind to the cis diol of the sugar on the antibody under physiological pH conditions. The artificially synthesized secondary antibody solves the problems of high production cost, difficult preservation and transportation and poor stability of biological secondary antibodies, and is suitable for a common ELISA process.
The invention provides an artificial secondary antibody taking graphene oxide as a carrier, which comprises the following raw materials in percentage by mass:
MES (2-morpholinoethanesulfonic acid): 0.02-0.19
EDC (1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride): 0.01-0.03
NHS (N-hydroxysuccinimide): 0.02-0.06
Graphene Oxide (GO): 0.01-0.03
Sodium cyanoborohydride: 0.77
Phenolphthalein or thymolphthalein: 0.02-0.06
2, 4-difluoro-3-formylphenylboronic acid or 2, 6-difluoro-4-formylphenylboronic acid or 2-formylpyridine-5-boronic acid pinacol ester: 0.02-0.05
BSA (bovine serum albumin): 0.001-0.004
Hexamethylenediamine or oxalyl-dihydrazide or polyoxyethylene diamine (MW 200, 400, 600, 800, 1000) or hydrazide-polyethylene glycol-hydrazide (MW 400, 600, 800, 1000, 2000): 0.02-0.05
pH12-pH14 lye (sodium hydroxide solution): 0.08-8
Ethanol: 0.02-0.08
Sodium dihydrogen phosphate: 0.11-0.22
Disodium hydrogen phosphate: 1.12-2.24
DMSO (dimethyl sulfoxide): 0.20-0.53
Water: 87.67-98.25
The sum of the mass percentages of the raw materials is 100.
The synthesis steps comprise: grafting a spacer on graphene oxide; boric acid functionalization is realized on graphene oxide; loading a pH sensitive color developing agent on the functional graphene oxide; covalent bonding is performed by utilizing boron affinity, and pH is adjusted to enable the color developing agent to develop color.
The synthetic method of the artificial secondary antibody with the graphene oxide as the carrier provided by the invention comprises the following steps:
1) Grafting hexamethylenediamine, oxalyl dihydrazide, polyoxyethylene diamine or hydrazide-polyethylene glycol-hydrazide spacer on graphene oxide through peptide forming reaction or ring opening reaction; specifically, GO is firstly dispersed in MES buffer solution by ultrasonic, EDC/NHS is then added for activation, and then the grafting agent is added for reaction;
2) And adding boric acid functional reagent: 2, 4-difluoro-3-formylphenylboric acid, 2, 6-difluoro-4-formylphenylboric acid and 2-formylpyridine-5-boric acid pinacol ester are grafted at the tail end of a spacing arm to be subjected to covalent bonding to synthesize an artificial secondary antibody;
3) The color-developing agent phenolphthalein or thymolphthalein is loaded on graphene oxide through hydrophobic action and pi-pi action, specifically, the phenolphthalein or thymolphthalein is dissolved in DMSO, and is added into PBS (phosphate buffer solution) dispersion liquid of GO, and the color-developing agent is adsorbed on GO through a simple solid-phase extraction process.
The invention provides an application of an artificial secondary antibody with graphene oxide as a carrier, which is applied to common ELISA: and (3) adding the synthesized functional GO dispersion liquid into a commercial ELISA process, covalently combining the synthesized functional GO dispersion liquid with cis-diol at the tail end of an antibody, washing, adding alkali liquor, dissociating a pH sensitive color-developing agent from GO into the solution for color development, and then measuring an OD (absorbance) value to quantitatively react with a substrate instead of enzyme.
The invention is to incubate boric acid and antibody Fc segment or glycoprotein sugar chain covalent bond under physiological pH, and to make the color-developing agent develop by adjusting pH, which can effectively replace biological secondary anti-labelling antibody and subsequent enzyme and substrate color reaction. The space resistance of boron affinity is effectively reduced through grafting the spacer arm, the pH value of boron affinity combination is reduced to a physiological condition through F-substituted boric acid or pyridine boric acid, the graphene oxide has a very large surface area, and the color development condition is optimized and the color development step is simplified through loading phenolphthalein or thymolphthalein.
Compared with the prior art, the invention has the following beneficial effects:
(1) The chemical material is adopted to replace biological reagent, so that the cost of laboratory test reagent is reduced;
(2) The steps are simplified, the experiment time is shortened, and the labor cost is reduced;
(3) The stability is greatly improved, and the transportation and storage cost is reduced.
Drawings
Fig. 1 is a schematic identification process of an artificial secondary antibody (functional graphene oxide).
Fig. 2 is a synthetic process of artificial secondary antibodies (functional graphene oxide).
FIG. 3 shows the result of the application of artificial secondary antibodies.
FIG. 4 is a transmission electron microscope image of an artificial secondary antibody.
Detailed Description
The invention will be further described with reference to the drawings and detailed description. The described embodiments are some, but not all, embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to fall within the scope of the invention.
Example 1
5-15 mg Graphene Oxide (GO) was dispersed in 25ml 2-morpholinoethanesulfonic acid (MES) buffer (ph=6), vigorously sonicated 2-3 h, and activated by gentle shaking 12 h at room temperature with the addition of EDC (15 mg) and NHS (30 mg). The activated GO was washed 3 times by centrifugation, dispersed in PBS, added with 10-25-mg hexamethylenediamine and shaken at room temperature for 12-h, washed by centrifugation, redispersed in absolute ethanol (1% sodium cyanoborohydride), added with 10-25-mg of 2, 6-difluoro-4-formylphenylboronic acid, shaken for 6-12-h, and further added with 100-200. Mu.L of 1% Bovine Serum Albumin (BSA) solution to inactivate 2 h to block the remaining active sites. Adding 100-250 μl of 5-40 mM phenolphthalein/DMSO solution, oscillating at room temperature for 6-12 h, centrifuging, washing for 3 times, dispersing in 5mL 20 mM PBS (pH 7.4), and storing.
Example 2
5-15 mg Graphene Oxide (GO) was dispersed in 25ml 2-morpholinoethanesulfonic acid (MES) buffer (ph=6), vigorously sonicated 2-3 h, and activated by gentle shaking 12 h at room temperature with the addition of EDC (15 mg) and NHS (30 mg). The activated GO was washed 3 times by centrifugation, dispersed in PBS, added with 10-25 of mg polyoxyethylene diamine (MW 600) and shaken at room temperature for 12-h, washed by centrifugation, re-dispersed in absolute ethanol (1% sodium cyanoborohydride), added with 10-25-mg of 2, 6-difluoro-4-formylphenylboronic acid, shaken for 6-12-h, and further added with 100-200. Mu.L of 1% Bovine Serum Albumin (BSA) solution to inactivate 2-h to block the remaining active sites. Adding 100-250 μl of 5-40 mM phenolphthalein/DMSO solution, oscillating at room temperature for 6-12 h, centrifuging, washing for 3 times, dispersing in 5mL 20 mM PBS (pH 7.4), and storing.
Example 3
5-15 mg Graphene Oxide (GO) was dispersed in 25ml 2-morpholinoethanesulfonic acid (MES) buffer (ph=6), vigorously sonicated 2-3 h, and activated by gentle shaking 12 h at room temperature with the addition of EDC (15 mg) and NHS (30 mg). The activated GO was washed 3 times by centrifugation, dispersed in PBS, added with 10-25 of mg polyoxyethylene diamine (MW 600) and shaken at room temperature for 12-h, washed by centrifugation, re-dispersed in absolute ethanol (1% sodium cyanoborohydride), added with 10-25-mg of 2, 4-difluoro-3-formylphenylboronic acid, shaken for 6-12-h, and further added with 100-200. Mu.L of 1% Bovine Serum Albumin (BSA) solution to inactivate 2-h to block the remaining active sites. Adding 10-100 μl of 5-30 mM thymolphthalein/DMSO solution, oscillating at room temperature for 6-12 h, centrifuging, washing for 3 times, dispersing in 5mL 20 mM PBS (pH 7.4), and storing.
Example 4
100 mu L of transferrin solution with the concentration of 5-80 ppm is added into a commercial transferrin ELISA kit 96-well plate, incubated for 2 h at 37 ℃ and washed 3 times, then 100 mu L of transferrin antibody solution is added, incubated for 2 h at 37 ℃ and washed 3 times, 100-300 mu L of the artificial secondary antibody prepared in example 2 is added, incubated for 30 min at 37 ℃ and washed 3 times, 100-300 mu L of alkaline solution (aqueous sodium hydroxide solution) with the pH of 12 is added for developing for 2 min, OD (absorbance) values are measured at 553 nm wavelengths, and the development results of transferrin with different concentrations are shown in FIG. 3. The artificial secondary antibodies are suitable for use in a general ELISA procedure.
Claims (3)
1. Application of artificial secondary antibody with graphene oxide as carrier in enzyme-linked immunosorbent assay (ELISA) process; the synthetic method of the artificial secondary antibody taking graphene oxide as a carrier comprises the following steps of:
1) Grafting hexamethylenediamine, oxalyl dihydrazide, polyoxyethylene diamine or hydrazide-polyethylene glycol-hydrazide spacer on graphene oxide through peptide forming reaction or ring opening reaction; specifically, graphene oxide is firstly dispersed in 2-morpholinoethanesulfonic acid buffer solution in an ultrasonic way, then 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride/N-hydroxysuccinimide is added for activation, and then the grafting agent is added for reaction with the graphene oxide;
2) And adding boric acid functional reagent: 2, 4-difluoro-3-formylphenylboric acid, 2, 6-difluoro-4-formylphenylboric acid and 2-formylpyridine-5-boric acid pinacol ester are grafted at the tail end of a spacing arm to be subjected to covalent bonding to synthesize an artificial secondary antibody;
3) Loading and enriching a color developing agent phenolphthalein or thymolphthalein on graphene oxide through hydrophobic action and pi-pi action, dissolving phenolphthalein or thymolphthalein in dimethyl sulfoxide, adding the phenolphthalein or thymolphthalein into phosphate buffer solution dispersion liquid of the graphene oxide, and enabling the color developing agent to be adsorbed on the graphene oxide through a simple solid phase extraction process;
the artificial secondary antibody taking graphene oxide as a carrier comprises the following raw materials in percentage by mass:
2-morpholinoethanesulfonic acid: 0.02-0.19
1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride: 0.01-0.03
N-hydroxysuccinimide: 0.02-0.06
Graphene oxide: 0.01-0.03
Sodium cyanoborohydride: 0.77
Phenolphthalein or thymolphthalein: 0.02-0.06
2, 4-difluoro-3-formylphenylboronic acid or 2, 6-difluoro-4-formylphenylboronic acid or 2-formylpyridine-5-boronic acid pinacol ester: 0.02-0.05
Bovine serum albumin: 0.001-0.004
Hexamethylenediamine or oxalyl dihydrazide or polyoxyethylene diamine or hydrazide-polyethylene glycol-hydrazide: 0.02-0.05
pH12-pH14 lye: 0.08-8
Ethanol: 0.02-0.08
Sodium dihydrogen phosphate: 0.11-0.22
Disodium hydrogen phosphate: 1.12-2.24
Dimethyl sulfoxide: 0.20-0.53
Water: 87.67-98.25
The sum of the mass percentages of the raw materials is 100.
2. The method according to claim 1, wherein the grafting agent is hexamethylenediamine.
3. The use according to claim 1, characterized in that the boric acid functional agent is: 2, 4-difluoro-3-formylphenylboronic acid, 2, 6-difluoro-4-formylphenylboronic acid.
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|---|---|---|---|---|
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| CN103212084A (en) * | 2003-11-13 | 2013-07-24 | 韩美科学株式会社 | IgG FC fragment for a drug carrier and method for the preparation thereof |
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| CN111579467A (en) * | 2020-06-01 | 2020-08-25 | 天津医科大学 | Bifunctional graphene oxide composite material and application thereof in detecting adherent cells |
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Patent Citations (4)
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|---|---|---|---|---|
| CN103212084A (en) * | 2003-11-13 | 2013-07-24 | 韩美科学株式会社 | IgG FC fragment for a drug carrier and method for the preparation thereof |
| WO2009066275A1 (en) * | 2007-11-22 | 2009-05-28 | Dublin City University | A method of immobilising biological molecules to a support and products thereof |
| CN104155357A (en) * | 2014-05-23 | 2014-11-19 | 济南大学 | Preparation method and application of three-dimensional cubic duct based mesoporous silica sensor |
| CN111579467A (en) * | 2020-06-01 | 2020-08-25 | 天津医科大学 | Bifunctional graphene oxide composite material and application thereof in detecting adherent cells |
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| Title |
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