CN110108656A - A kind of method of the fixed uricase detection uric acid of mesoporous organosilicon hollow nanospheres - Google Patents
A kind of method of the fixed uricase detection uric acid of mesoporous organosilicon hollow nanospheres Download PDFInfo
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- 229940116269 uric acid Drugs 0.000 title claims abstract description 98
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 title claims abstract description 93
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 title claims abstract description 93
- 108010092464 Urate Oxidase Proteins 0.000 title claims abstract description 82
- 239000002077 nanosphere Substances 0.000 title claims abstract description 73
- 238000001514 detection method Methods 0.000 title claims abstract description 56
- 238000000034 method Methods 0.000 title claims abstract description 33
- 230000015572 biosynthetic process Effects 0.000 claims abstract description 21
- 238000003786 synthesis reaction Methods 0.000 claims abstract description 21
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 13
- 239000000377 silicon dioxide Substances 0.000 claims abstract description 11
- 229910052681 coesite Inorganic materials 0.000 claims abstract description 9
- 229910052906 cristobalite Inorganic materials 0.000 claims abstract description 9
- 229910052682 stishovite Inorganic materials 0.000 claims abstract description 9
- 229910052905 tridymite Inorganic materials 0.000 claims abstract description 9
- 238000002835 absorbance Methods 0.000 claims description 31
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 28
- 108090000790 Enzymes Proteins 0.000 claims description 25
- 102000004190 Enzymes Human genes 0.000 claims description 25
- 239000000243 solution Substances 0.000 claims description 24
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 18
- 235000019441 ethanol Nutrition 0.000 claims description 13
- 239000000203 mixture Substances 0.000 claims description 11
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 11
- 239000012498 ultrapure water Substances 0.000 claims description 11
- 239000004471 Glycine Substances 0.000 claims description 9
- 239000000872 buffer Substances 0.000 claims description 8
- 238000003756 stirring Methods 0.000 claims description 8
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 7
- 239000000908 ammonium hydroxide Substances 0.000 claims description 7
- 238000005119 centrifugation Methods 0.000 claims description 7
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 5
- 239000011259 mixed solution Substances 0.000 claims description 5
- FOQJQXVUMYLJSU-UHFFFAOYSA-N triethoxy(1-triethoxysilylethyl)silane Chemical compound CCO[Si](OCC)(OCC)C(C)[Si](OCC)(OCC)OCC FOQJQXVUMYLJSU-UHFFFAOYSA-N 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 4
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 claims description 3
- BOTDANWDWHJENH-UHFFFAOYSA-N Tetraethyl orthosilicate Chemical compound CCO[Si](OCC)(OCC)OCC BOTDANWDWHJENH-UHFFFAOYSA-N 0.000 claims description 3
- 238000000227 grinding Methods 0.000 claims description 3
- 238000004108 freeze drying Methods 0.000 claims description 2
- 239000002244 precipitate Substances 0.000 claims description 2
- 238000006467 substitution reaction Methods 0.000 claims description 2
- 238000011010 flushing procedure Methods 0.000 claims 1
- 210000002966 serum Anatomy 0.000 abstract description 30
- 230000008569 process Effects 0.000 abstract description 7
- 238000002474 experimental method Methods 0.000 abstract description 5
- 230000007797 corrosion Effects 0.000 abstract description 3
- 238000005260 corrosion Methods 0.000 abstract description 3
- 229940088598 enzyme Drugs 0.000 description 24
- 230000000694 effects Effects 0.000 description 23
- 239000000523 sample Substances 0.000 description 17
- 239000007788 liquid Substances 0.000 description 11
- 238000006243 chemical reaction Methods 0.000 description 9
- 239000000463 material Substances 0.000 description 9
- 239000006228 supernatant Substances 0.000 description 7
- 241000282414 Homo sapiens Species 0.000 description 6
- 239000011805 ball Substances 0.000 description 5
- 239000012876 carrier material Substances 0.000 description 5
- POJWUDADGALRAB-UHFFFAOYSA-N allantoin Chemical compound NC(=O)NC1NC(=O)NC1=O POJWUDADGALRAB-UHFFFAOYSA-N 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012895 dilution Substances 0.000 description 4
- 230000002255 enzymatic effect Effects 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 239000002086 nanomaterial Substances 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 230000005540 biological transmission Effects 0.000 description 3
- 238000002329 infrared spectrum Methods 0.000 description 3
- 239000000696 magnetic material Substances 0.000 description 3
- 239000011148 porous material Substances 0.000 description 3
- 229910052710 silicon Inorganic materials 0.000 description 3
- 239000010703 silicon Substances 0.000 description 3
- 239000004094 surface-active agent Substances 0.000 description 3
- POJWUDADGALRAB-PVQJCKRUSA-N Allantoin Natural products NC(=O)N[C@@H]1NC(=O)NC1=O POJWUDADGALRAB-PVQJCKRUSA-N 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- GWEVSGVZZGPLCZ-UHFFFAOYSA-N Titan oxide Chemical compound O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 description 2
- 229960000458 allantoin Drugs 0.000 description 2
- 230000004075 alteration Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000003431 cross linking reagent Substances 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000007306 functionalization reaction Methods 0.000 description 2
- 230000035484 reaction time Effects 0.000 description 2
- 238000004064 recycling Methods 0.000 description 2
- 238000012552 review Methods 0.000 description 2
- 239000013049 sediment Substances 0.000 description 2
- JIOGKDWMNMIDEY-UHFFFAOYSA-N triethoxy-(2-triethoxysilylphenyl)silane Chemical compound CCO[Si](OCC)(OCC)C1=CC=CC=C1[Si](OCC)(OCC)OCC JIOGKDWMNMIDEY-UHFFFAOYSA-N 0.000 description 2
- -1 triethoxysilyl Chemical group 0.000 description 2
- 238000002604 ultrasonography Methods 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 238000004566 IR spectroscopy Methods 0.000 description 1
- 108010093096 Immobilized Enzymes Proteins 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 229910018540 Si C Inorganic materials 0.000 description 1
- OBNDGIHQAIXEAO-UHFFFAOYSA-N [O].[Si] Chemical compound [O].[Si] OBNDGIHQAIXEAO-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000012496 blank sample Substances 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 238000001354 calcination Methods 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 244000145845 chattering Species 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 238000004945 emulsification Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000003628 erosive effect Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000003862 health status Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000004750 isotope dilution mass spectroscopy Methods 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 239000013337 mesoporous metal-organic framework Substances 0.000 description 1
- 239000012621 metal-organic framework Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
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- 238000012544 monitoring process Methods 0.000 description 1
- 239000011807 nanoball Substances 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 238000000643 oven drying Methods 0.000 description 1
- 230000020477 pH reduction Effects 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 238000006068 polycondensation reaction Methods 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
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- 235000009566 rice Nutrition 0.000 description 1
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- 238000001338 self-assembly Methods 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 229910010271 silicon carbide Inorganic materials 0.000 description 1
- 238000003980 solgel method Methods 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
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- 230000002194 synthesizing effect Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- KENDGHJJHKCUNB-UHFFFAOYSA-N triethoxy-[4-(4-triethoxysilylphenyl)phenyl]silane Chemical group C1=CC([Si](OCC)(OCC)OCC)=CC=C1C1=CC=C([Si](OCC)(OCC)OCC)C=C1 KENDGHJJHKCUNB-UHFFFAOYSA-N 0.000 description 1
- 230000002485 urinary effect Effects 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
Abstract
A kind of method of the fixed uricase detection uric acid of mesoporous organosilicon hollow nanospheres, belongs to uric acid detection technique field.This experiment is with monodisperse SiO2Nanosphere prepares mesoporous organosilicon hollow nanospheres as hard template, using one step growth induced corrosion method.This method is simple to operation, it is desirable that not high, synthesis process is highly controllable, and the mesoporous organosilicon nanosphere of the hollow structure with different function group can be synthesized by using different presomas.The mesoporous organosilicon hollow nanospheres duct of synthesis is orderly, and aperture is uniform, and hollow cavity is contained in inside.Serum uric acid is detected using the mesoporous organosilicon hollow nanospheres fixed uricase after synthesis, a kind of method for establishing fixed uricase detection uric acid of mesoporous organosilicon hollow nanospheres has application prospect in the quick context of detection of serum uric acid.
Description
Technical field
The invention belongs to uric acid detection technique fields, and in particular to a kind of fixed uricase of mesoporous organosilicon hollow nanospheres
The method for detecting uric acid.
Background technique
The health status of the content and many systems of human body of internal serum uric acid is closely bound up, generation with a variety of diseases and
It is in progress all relevant.Therefore, the content of detection in time and monitoring serum uric acid has weight to the prevention and treatment of the health and disease of human body
Want meaning.
The common method of detection uric acid mainly has high performance liquid chromatography, isotope dilution mass spectrometry, enzyme process etc. at present, preceding
Two kinds of detection methods the disadvantages of there are complicated for operation, expensive equipments, long detection cycle, it is unsuitable for the quick detection of uric acid.And enzyme
Method detects uric acid and generates allantoin by uric acid enzymatic, and the product of generation is directly proportional to concentration of substrate, by measuring allantoin
Absorbance value, calculate the concentration of uric acid, enzyme process detection have many advantages, such as it is specific it is good, accuracy is high, but uricase is to reaction
Condition requires height, can not recycle the method for continuing with [Wang Jing detection blood uric acid and compare and be in progress [J] diabetes clinic,
2014 (8): 362-263], so using enzyme immobilization technology, using the fixed uricase of mesoporous organosilicon hollow nanospheres to blood
Clear uric acid is measured.
The carrier material of fixed uricase mainly has high molecular material, magnetic material, nano material etc. at present.Macromolecule carries
Body mechanical strength is low, and microorganism can be easy to decompose carrier material, and fixed uricase is caused to fall off, and causes to carry
Body-uricase can not repeatedly use, and polymer carrier service life is short.And magnetic carrier material is usually to pass through
The fixed uricase of the mode of crosslinking, needs to clean crosslinking agent, the fixation to enzyme be not it is highly stable, crosslinking agent may also influence
The activity of enzyme.Nano material carrier is more but usually used in papain, fatty enzyme immobilizatio application study in recent years
Be nano magnetic material, this needs modifying agent to modify nano magnetic material, modification need condition it is quicker
Sense, and be easy to impact [Dong Y.Y.Recent Progress of Materials Used as to enzymatic activity
Enzyme Immobilization Carriers[J].Journal of Shanghai Institute of
Technology,2013(13):295-298.】。
Compared with these traditional materials, mesoporous organosilicon hollow nanospheres have very big advantage in terms of immobilized enzyme.
The porous nanometer material of hollow structure is because of its low-density, high-specific surface area, permeable shell and can functionalization hollow cavity and
By the extensive concern of researcher, the core of functionalization can move freely that its active site is made to become more to hold in cavity
It is accessible, at the same the chemical composition of core and shell can multiplicity change, therefore the porous material of these nanostructures by
It is widely applied to different fields [Lee J., Farha O., Roberts J.Metal-organic framework
materials as catalysts[J].Chemical Society Reviews,2009(23):1450-1453.】【Xuan
W.,Zhu C.,Liu Y.Mesoporous metal–organic framework materials[J].Chemical
Society Reviews,2012(41):1677-1684.].The mesoporous organosilicon (HPMO) of hollow structure can be by nanostructure
Characteristic and the characteristic of PMO combine, great application potential can be shown, be the project for being worth further investigation,
However, up to the present, it is actually rare to the report of HPMO.
Under normal circumstances, hollow structure PMO[Yang J., Lind J.U. are mainly synthesized using soft template and hard template method,
Trogler W.C.Synthesis of Hollow Silica and Titania Nanospheres[J].Chemical
Materials, 2008 (20): 2875-2879.], the mesoporous organosilicon porous material of soft template method synthesis hollow structure is mainly
By surfactant, the self assembly of presoma and hydrolytie polycondensation, this method makes surfactant in solution-air or liquid-liquid interface
Emulsification, it is difficult to ensure that the regulation of the homogeneity of template and scantling.And hard template rule can overcome more than disadvantage, it is right
It can be controlled in aperture control and parameter change proper.Hard template method synthesizes the mesoporous organosilicon porous material selection of hollow structure
Uniform monodisperse nanoparticle is as hard template, using sol-gel method in hard template Surface Creation mesoporous organosilicon shell, so
Pass through calcining or other acid processing removal cores and surfactant afterwards.However, these methods are directed to a series of cladding-corruption
The complicated process such as erosion, the process of these multisteps needs to put into more energy and times, and often requires to use toxic
Reagent, meanwhile, the mesopore orbit of obtained PMO hollow shell be it is unordered, direction is arbitrary, this is limited to a certain extent
The infiltration of guest molecule, therefore, the simple and effective step strategy synthesis of development one have orderly vertical mesopore orbit
HPMO is still a very big challenge.
The fixed usual adsorbance of uricase of traditional carrier material is small, and secure bond is insecure, and is easy to inhibit uricase
Activity, be unfavorable for the reuse of uricase detection uric acid, therefore fixed amount is big, does not destroy the carrier material of uric acid enzymatic structure
Always study the hot spot of concern.Therefore orderly controllable mesoporous the purpose of the present invention is synthesizing there is hollow cavity and duct
Organosilicon hollow nanometer material is fixed uricase with it and is detected to serum uric acid.
Summary of the invention
The present invention is with monodisperse SiO2As hard template, prepared using one step growth induced corrosion method mesoporous is had nanosphere
Machine silicon hollow nanospheres.This method is simple to operation, it is desirable that not high, synthesis process is highly controllable, and by using different
Presoma, such as bis- (triethoxysilyl) ethane (BTEE) of 1,2-, bis- (triethoxysilyl) benzene (BTEB) of Isosorbide-5-Nitrae-
With bis- (triethoxysilyl) biphenyl (BTEBP) of 4,4-, Jie of the hollow structure with different function group can be synthesized
Hole organic silicon nano ball.The mesoporous organosilicon hollow nanospheres duct of synthesis is orderly, and aperture is uniform, and hollow cavity is contained in inside.
The present invention utilizes organosilan 1, and bis- (triethoxysilyl) ethane of 2- are as precursor synthesis hollow structure
Mesoporous organosilicon hollow nanospheres.The content of present invention mainly includes that synthesis duct is orderly, and aperture is uniform, and hollow cavity is contained in inside
Mesoporous organosilicon hollow nanospheres, using scanning electron microscope, transmission electron microscope, infrared spectroscopy is hollow to the mesoporous organosilicon of synthesis receives
Rice ball is characterized, and is detected, is built to serum uric acid using the fixed uricase of mesoporous organosilicon hollow nanospheres after synthesis
A kind of method for founding fixed uricase detection uric acid of mesoporous organosilicon hollow nanospheres.
The present invention has very strong adsorptivity and highly controllable duct, Ke Yiti using mesoporous organosilicon hollow nanospheres
The fixed amount of high lithemia enzyme keeps the activity of uricase, and the recycling of uricase can be improved in the uricase detection uric acid after fixing
Utilization rate shortens the time of uric acid detection, expands the detection range of uric acid, before the quick context of detection of serum uric acid has application
Scape.
A kind of method of the fixed uricase detection uric acid of mesoporous organosilicon hollow nanospheres of the present invention, step is such as
Under:
(1) hard template monodisperse SiO2The synthesis of nanosphere: it is ultrapure that 3mL ethyl orthosilicate is added to 37mL ethyl alcohol, 5mL
In the mixture of water and 1.6mL ammonium hydroxide (mass fraction 28%), cleans and dry after stirring centrifugation, obtain monodisperse SiO2Nanometer
Ball；
(2) synthesis of mesoporous organosilicon hollow nanospheres: the monodisperse SiO that 100mg step (1) is synthesized2Nanosphere is logical
Ultrasonic disperse is crossed in the mixture of 11mL ethyl alcohol and 22mL ultrapure water, then by 120mg cetyl trimethylammonium bromide and 2mL
Ammonium hydroxide (mass fraction 28%), 0.22mL bis- (triethoxysilyl) ethane of 1,2- be added in above-mentioned mixture,
Kettle is filled after stirring 2h, then stands for 24 hours, is cooled to room temperature at 100 DEG C, is centrifugated, and with water and ethanol washing, at 80 DEG C
It is extracted after drying with 65mL and 2mL concentrated hydrochloric acid (mass fraction 36%), it is obtained by drying after centrifuge washing to arrive mesoporous organosilicon
Hollow nanosphere carrier, grinding are stand-by；
(3) fixed uricase: it is dense that 10mL is added in the mesoporous organosilicon hollow nanosphere carrier for taking 10mg step (2) to grind
Degree is the uric acid enzyme solution of 0.2mg/mL, stirs 8h under 4 DEG C of environment, and taking precipitate is lyophilized to get carrier-uricase is arrived after centrifugation；
(4) carrier-uricase for the quality 3mg that 7 parts of steps (3) obtain is added separately to the sweet ammonia of 2mL, pH=8.5
In the mixed solution of acid buffer and 0.5mL ultrapure water, then be separately added into 0.5mL, concentration be respectively 0.001mg/mL,
The uric acid titer of 0.01mg/mL, 0.05mg/mL, 0.1mg/mL, 0.5mg/mL, 1.0mg/mL, 1.5mg/mL, 40 DEG C of water-baths
20min is reacted, then detects absorbance value of the mixed liquor at 290nm respectively using ultraviolet specrophotometer；With uric acid standard
The concentration of liquid is abscissa, makes standard curve by ordinate of the absorbance value at the 290nm measured；
(5) by carrier-uricase that 3mg step (3) obtains be added to 2mL, pH=8.5 glycine buffer and
In the mixed solution of 0.5mL ultrapure water, the uric acid solution of 0.5mL, unknown concentration are added, 40 DEG C of water-bath 20min, then
Use absorbance value of the UV spectrophotometer measuring mixed liquor at 290nm；Absorbance value substitution step (4) is obtained
In standard curve, the concentration of the uric acid solution is calculated, realizes the detection to uric acid.
The present invention is a kind of novel mesoporous organosilicon hollow nanospheres synthesis and its fixed uricase detection uric acid, spy
Sign is:
(1) each ball of mesoporous organosilicon hollow nanospheres is hollow, average-size 380nm, and each hollow ball
It all include a uniform 75nm Et-PMO shell, hollow cavity 230nm, size is exactly equal to SiO2The partial size of nanosphere.
(2) compared with free uricase, the uricase activity after fixing still is able to maintain 95% or more.Free uricase is most
Thermophilic degree is 30 DEG C, optimal pH 10, and carrier-uricase optimum temperature and pH are respectively 40 DEG C and 8.5.Free uricase
More stable between pH=8-10, meta-alkalescence, carrier-uricase pH stability boundary becomes more extensive, in pH=7-
It is more stable between 11.
(3) the fixed uricase of mesoporous organosilicon hollow nanospheres detects serum uric acid, and optimum reacting time is 20 minutes, with
The reaction time of general uricase detection uric acid is many compared to shortening.When carrier-uricase of 3mg is added, reaction can be into
Completely, the amount of mesoporous organosilicon Hollow nanosphere material used and uricase is all little, can obtain good detection effect for row.
It is linear good in the range of 0.01mg/mL~1.0mg/mL, R2=0.9982.Mesoporous organosilicon hollow nanospheres fix uric acid
Enzyme detects serum uric acid, and carrier uricase remains to reuse after centrifugation, identical as detection uric acid standard items effect,
After repeating detection serum uric acid 40 times, the activity of carrier uricase is just lost.
Detailed description of the invention
Fig. 1 (A), (B) are monodisperse SiO2The scanning electron microscope (SEM) photograph of nanosphere and mesoporous organosilicon hollow nanospheres, Fig. 1 (C)
For transmission electron microscope picture.Fig. 1 (A) is the monodisperse SiO that embodiment 1 synthesizes2The scanning electron microscope (SEM) photograph of nanosphere, can be with from Fig. 1 (A)
The monodisperse SiO of synthesis visible in detail2The size uniformity of nanosphere, topographic profile is good, and average-size is on the left side 230nm
It is right.Fig. 1 (B) is the scanning electron microscope (SEM) photograph for the mesoporous organosilicon hollow nanospheres that embodiment 2 synthesizes, we can from Fig. 1 (B)
Mesoporous organosilicon hollow nanospheres to synthesis are well dispersed, and pattern understands.Fig. 1 (C) is the mesoporous organosilicon that embodiment 2 synthesizes
The transmission electron microscope picture of hollow nanospheres, from Fig. 1 (C) it can be seen that each mesoporous organosilicon hollow nanospheres of synthesis are hollow
, the size of internal cavities is 230nm, with monodisperse SiO2The size of nanosphere is consistent.Each ball is uniform containing one,
Organosilan shell having a size of 75nm, the average-size of mesoporous organosilicon hollow nanospheres are 380nm.
Fig. 2 is infrared spectrogram of the mesoporous organosilicon hollow nanospheres of the synthesis of embodiment 2 before and after fixed uricase, a
For the infrared spectrum curve of uricase, b is the infrared spectrum curve of mesoporous organosilicon hollow nanospheres, and c is in mesoporous organosilicon
The infrared spectrum curve of the fixed uricase of empty nanosphere.It can be observed from fig. 2 that mesoporous organosilicon hollow nanospheres and fixed uric acid
Mesoporous organosilicon hollow nanospheres after enzyme are in 1410cm-1The absorption band for thering is a c h bond to vibrate, in 1160cm-1There is one
The absorption band of Si-C key chattering, this shows that the organic (- Et-) silicon of bridged group is present in sample.In 2950cm-1Neighbouring uricase
There is an absorption band with the mesoporous organosilicon hollow nanospheres after fixed uricase, shows that uricase is successfully fixed
Onto mesoporous organosilicon hollow nanospheres.
Fig. 3 is the standard curve of 3 uricase absorbance of embodiment, can calculate uricase according to the absorbance standard curve
Fixed amount.By can be seen that the concentration (abscissa) of uricase is directly proportional to absorbance value (ordinate) on figure, curvilinear equation y
=1.0018x (R2=0.9978).
Fig. 4 is 6 mesoporous organosilicon hollow nanospheres of embodiment fixed uricase (carrier-uricase) detection uric acid titer
Standard curve, abscissa be uric acid titer concentration, ordinate be absorbance value of the supernatant at 290nm.It can by Fig. 4
To see that this law detection uric acid is linear good in the range of 0.001mg/mL~1.5mg/mL, linear equation y=
0.8246x-0.0026(R2=0.9995).
Fig. 5 is 7 carriers of embodiment-uricase detection serum uric acid canonical plotting, and abscissa is the dense of serum uric acid
Degree, ordinate are absorbance value of the supernatant at 290nm.By Fig. 5, it can be seen that, carrier-uricase detection serum uric acid exists
Linear good in the range of 0.001~1.0mg/mL, linear equation is y=0.7714x+0.0133 (R2=0.9982).Human body
The normal concentration range of serum uric acid is 0.015~0.07mg/mL, it can be seen that the concentration range packet of this law detection serum uric acid
The concentration of the serum uric acid containing human normal.
Specific embodiment
The present invention is further illustrated combined with specific embodiments below, but protection scope of the present invention is not limited to
This.
On the one hand the present invention provides the synthesis of mesoporous organosilicon hollow nanospheres, we are according to silica chemistry and silicon oxygen
The similitude of alkanisation has developed a kind of one step growth induced corrosion method preparation of organosilan guidance with orderly vertical mesoporous
The mesoporous organosilicon hollow nanospheres in duct.
Embodiment 1
(1) 37mL ethyl alcohol and 5mL ultrapure water are taken with graduated cylinder correct amount, poured into 100mL beaker.
(2) ammonium hydroxide (mass fraction 28%) of 1.6mL is added, is then rapidly added the ethyl orthosilicate of 3mL.
(3) supercentrifuge 12000rpm is used to be centrifuged 3min after stirring 2h at room temperature.
(4) cleaned twice with ethyl alcohol again after being cleaned twice with ultrapure water, after centrifugation room temperature naturally dry to get to quality about
The monodisperse SiO of 300mg2Nanosphere.
Embodiment 2
(1) 22mL ultrapure water and 11mL ethyl alcohol are accurately measured in 100mL beaker.
(2) the 100mg monodisperse SiO that Example 1 synthesizes2Nanosphere is added thereto, mixed liquor ultrasound 30min.
(3) 120mg cetyl trimethylammonium bromide and 2mL ammonium hydroxide (mass fraction 28%) are added, is stirred at room temperature
Bis- (triethoxysilyl) second of 1,2- of 0.22mL are added dropwise after being completely dissolved to cetyl trimethylammonium bromide
Alkane continues to stir 2h at room temperature.
(3) said mixture is fitted into reaction kettle, reaction kettle is stood for 24 hours in 100 DEG C of baking ovens, it is cold takes out reaction kettle
But mixture 12000rpm in kettle is centrifuged 3min to after room temperature.
(4) it is respectively cleaned twice with ultrapure water and ethyl alcohol, puts it into 80 DEG C of baking ovens and dry.
(5) sample after taking out drying is added a little ethyl alcohol and carries out ultrasound, and 65mL ethyl alcohol and 2mL concentrated hydrochloric acid are added later
(mass fraction 36%) is extracted, and is then centrifuged mixture, and 12000rpm is centrifuged 3min.
(6) cleaned after 2 times with ethyl alcohol be put into baking oven drying, take out drying after intermediary hole organosilicon sky nanosphere grinding to
With.
Embodiment 3
(1) the uricase stoste for taking 1mg/mL, with the dilution of the glycine buffer of pH=8.5 prepare 0.1mg/mL,
The uric acid enzyme dilution of 0.2mg/mL, 0.3mg/mL, 0.4mg/mL, 0.5mg/mL measure uric acid enzyme dilution at 293nm
Absorbance establishes the standard curve (such as Fig. 3) of uricase absorbance, to calculate uricase fixed amount.
(2) take 2mL concentration be 1mg/mL uricase stoste, with pH be 8.5 glycine buffer dilute 5 times to get
The uric acid enzyme solution for being 0.2mg/mL to 10mL concentration.The 10mg mesoporous organosilicon hollow nanospheres synthesized are added thereto, 4 DEG C are stirred
Mix 8h.
(3) it is centrifuged 5min with supercentrifuge 12000rpm, takes supernatant to survey its absorbance at 293nm, according to step
Suddenly the standard curve (Fig. 3) of (1) calculates the concentration of uricase in supernatant, is 0.0372mg/mL；And then calculate consolidating for uricase
Quantitative, uricase fixed amount calculation formula is as follows:
L (mg/g)=(C0-Ct) V/m=(0.2mg/mL-0.0172mg/mL) 10mL/10mg=182.8mg/g
L is uricase fixed amount, C0It is the concentration of fixed preceding uricase, CtFor the concentration of uricase in supernatant after fixation,
V is that the volume 10mL, m of solution are the quality of support samples.
(4) carrier of lower layer-uricase sediment is put into vacuum freeze drier freeze-drying 12h, and 4 DEG C of refrigerators are put into after taking-up
In save backup.
Embodiment 4: uricase activity detection
(1) glycine buffer that 1mL, pH=8.5 is added in the carrier for taking the embodiment 4 of 1mg to obtain-uricase is made
1mg/mL carrier-uricase solution dilutes 4 times, i.e. carrier-uric acid enzyme solution concentration is 0.25mg/mL.
(2) 0.5mL ultrapure water and the uric acid titer of 2mL, 0.05mg/mL are taken, in 40 DEG C of water-bath 5min after mixing.
(3) carrier-uric acid enzyme solution is reacted with the mixed liquor of step (2) after 0.5mL step (1) dilution is added in experiment tube
The KOH termination reaction of 0.2mL, mass fraction 2% is added after 5min, surveys solution at 293nm using ultraviolet specrophotometer
Absorbance value.
(4) KOH of 0.2mL, mass fraction 2% are first added in the blank tube of control, adds 0.5mL step (1)
Carrier-uric acid enzyme solution, the absorbance value of solution is surveyed using ultraviolet specrophotometer at 293nm.
Uricase activity detection formula is as follows:
UmL-1=(ODblank-ODtest)Vtdf/12.04bVst
Umg-1=(U mL-1) 1/C=9.2U/mg
ODblank: the absorbance value of blank tube solution, measuring it is 0.989；ODtest: the absorbance value of experiment tube solution,
Measuring it is 0.00032；Vt: overall solution volume 3.2mL；Vs: sample solution volume 0.5mL；T: reaction time 5min；Df: dilute
Releasing multiple is 4 times；B: cuvette aperture 1cm, 12.04: uric acid titer absorptivity, C: the concentration of step (1) enzyme solution sample
For 0.0457mg/mL.
(5) activity for the free uricase that this experiment purchase uses is 10U/mg, and mesoporous organosilicon hollow nanospheres are fixed
The activity of uricase afterwards is 9.2U/mg, and activity maintains 92% compared with free uricase, it can be seen that, mesoporous organosilicon
Hollow nanospheres will not destroy the structure of uricase, cannot keep uricase activity.
Embodiment 5: the comparison of the fixed front and back zymologic property of uricase
We fix optimal reactive temperature, the optimal pH, pH stability, thermal stability of front and back to uricase in the present invention
It is compared, more objectively to observe the zymologic property of the fixed uricase of mesoporous organosilicon hollow nanospheres.Concrete operations
Steps are as follows:
(1) by 0.1mL concentration be 0.5mg/mL uric acid enzyme solution and 0.1mL concentration be 0.5mg/mL carrier-uricase
Liquid is respectively put into the glycine buffer that pH is 6.0,6.5,7.0,7.5,8.0,8.5,9.0,9.5,10,10.5,11, is passed through
Detection activity is to verify the optimal pH that mesoporous organosilicon hollow nanospheres fix front and back uricase.The optimal pH of uricase before fixed
It is 10, the optimal pH of uricase is 8.5 after fixing.
(2) respectively by 0.5mL concentration be 0.5mg/mL uricase enzyme solution and carrier-uric acid enzyme solution be put into 0 DEG C, 10 DEG C,
It is incubated in the water-bath of 20 DEG C, 30 DEG C, 40 DEG C, 50 DEG C, 60 DEG C different temperatures, by calculating enzymatic activity to verify in mesoporous organosilicon
The optimum temperature of the fixed front and back uricase of empty nanosphere.The optimum temperature of uricase is 30 DEG C before fixed, uricase after fixing
Optimum temperature is 40 DEG C.
(3) by 0.1mL concentration be 0.5mg/mL carrier-uric acid enzyme solution and uric acid enzyme solution be put into pH be, 4,5,6,7,8,
9, in 10,11,12 glycine buffer, pH Detection of Stability is carried out by measurement enzyme activity.Free uricase in pH=8-
10 range is more stable, and uricase is more stable within the scope of pH=7-11 after fixing.
(4) uricase after the free uricase of measurement and mesoporous organosilicon hollow nanospheres are fixed is in 40 DEG C, 50 DEG C, 60
DEG C, the thermal stability at a temperature of 70 DEG C.When temperature reaches 70 DEG C, the activity residue for the uricase that dissociates works as temperature less than 10%
When degree reaches 60 DEG C, the decline rapidly of the activity for the uricase that dissociates.When temperature reaches 70 DEG C, it is fixed after uricase activity still
40% or more is kept, even if reaction temperature reaches 60 DEG C, activity decline is also unobvious.
Embodiment 6: uric acid titer detection
(1) to 0.001mg/mL, 0.01mg/mL, 0.05mg/mL, 0.1mg/mL, 0.5mg/mL, 1.0mg/mL, 1.5mg/
The uric acid titer of mL concentration is detected.
(2) by 7 parts be carrier-uricase of 3mg be added separately to the pH=8.5 of 2mL glycine buffer and
In the mixed solution of the ultrapure water of 0.5mL, be finally separately added into 0.5mL, in the present embodiment step (1) various concentration uric acid mark
Quasi- liquid sample, 40 DEG C of water-bath 20min.
(3) it is centrifuged 4min with supercentrifuge 12000rpm after reaction, is mixed using UV spectrophotometer measuring
Absorbance value at liquid 290nm.
(4) abscissa is done with the concentration of uric acid titer sample, is made using measuring the absorbance value at 290nm as ordinate
Standard curve determines the range of linearity of uric acid detection.
(5) carrier-uricase solution of lower sediment is rinsed well, is continued uric acid detection, is made by 30 repetitions
With rear, activity still 70% or more residue, the results are shown in Table 1:
Table 1: carrier-uricase repeats detection uric acid titer residual activity
Embodiment 7
(1) blood serum sample comes from healthy volunteer, and serum is stored in subzero 20 DEG C of refrigerators, needed before use 3000rpm from
Heart 15min, takes supernatant to use.
(2) blood serum sample for taking 0.4mL, being separately added into 0.1mL concentration is 0.01mg/mL, 0.02mg/mL, 0.04mg/
The uric acid titer of mL, 0.08mg/mL, 0.1mg/mL, 0.2mg/mL, 0.4mg/mL, 0.8mg/mL, 1.0mg/mL, according to table 2
It is reacted, using the concentration of uric acid titer sample as abscissa, makes mark as ordinate to measure the absorbance value at 290nm
Directrix curve determines the range of linearity of serum uric acid detection.
(3) it takes the blood serum sample of 0.4mL to be added in developmental tube and control tube respectively, is then by 0.1mL concentration respectively
0.01mg/mL uric acid titer is added to developmental tube and standard pipe, is reacted according to table 2, after reaction, 12000rpm from
The absorbance of supernatant, sample absorbance value=control tube absorbance value+standard pipe absorbance value-examination are surveyed at the heart 4min, 290nm
Test pipe absorbance value.Blood serum sample uric acid concentration to be detected is calculated by the standard curve that step (2) are established, calculates and knows to be somebody's turn to do
The uric acid concentration of part blood serum sample is 0.042mg/mL, in normal concentration 0.015~0.07mg/mL range of human serum uric acid
It is interior.
Table 2: serum uric acid detection process
(4) it is continued to use after carrier-uricase centrifugation recycling, determines that carrier-uricase detection uric acid is reused
Number, repeat serum uric acid detect 40 times after, activity just disappearance, result such as table 3:
Table 3: carrier-uricase repeats detection serum uric acid residual activity
Methodology validation:
(1) after to deduct blank value, concentration value corresponding to 3 times of noises is the minimum detection limit of this experimental method.Most
Low detection limits C=3 × 0.00116/0.8824=0.0039mg/mL.
(2) blood serum sample for measuring 0.4mL is separately added into the uric acid titer of various concentration, by this law to serum uric acid into
The result of row detection and the result of free urinary acidification detection uric acid are for statistical analysis, are examined and are carried out using the T of independent sample
Analysis, t=0.007, df=16, p=0.995, p > 0.05, so it is believed that mesoporous organosilicon hollow nanospheres fix uricase
The testing result of detection uric acid and free uricase does not have difference.
(3) accuracy of this law is indicated with recovery test.Use blood serum sample that known concentration is 0.03mg/mL as
Blank sample, the uric acid titer of high, normal, basic three concentration of addition are respectively 0.1mg/mL, 0.05mg/mL, 0.01mg/mL,
According to established uric acid detection method detection assay liquid absorbance value at 290nm, is calculated and surveyed according to absorbance standard curve
Determine liquid concentration, the sample detection of each concentration 3 times is averaged.Measurement liquid mean concentration be respectively 0.1262mg/mL,
0.0781mg/mL, 0.03962mg/mL, the rate of recovery=(measurement liquid average measurement-raw sample liquid hold-up is average)/additional amount ×
100%, each concentration rate of recovery is respectively 96.2%, 94.7% and 93.4%, and the rate of recovery is 93.4%~96.2%, it is seen that this law
Accuracy is good.
(4) the uric acid titer for measuring 1mg/mL, is 0.01mg/ with 5% bovine serum albumin(BSA) matrix liquid compound concentration
The uric acid titer of mL, 0.1mg/mL, 1mg/mL, in a few days aberration rate is 2.52%~3.05%, and aberration rate is in the daytime
5.20%~6.87%.
Claims (3)
1. a kind of method of the fixed uricase detection uric acid of mesoporous organosilicon hollow nanospheres, its step are as follows:
(1) synthesis of mesoporous organosilicon hollow nanospheres: by 100mg monodisperse SiO2Nanosphere is by ultrasonic disperse in 11mL second
In the mixture of pure and mild 22mL ultrapure water, then by the 1,2- of 120mg cetyl trimethylammonium bromide and 2mL ammonium hydroxide, 0.22mL
Bis- (triethoxysilyl) ethane are added in above-mentioned mixture, are filled kettle after stirring 2h, are then stood at 100 DEG C
For 24 hours, be cooled to room temperature, be centrifugated, and use water and ethanol washing, it is dry at 80 DEG C after with 65mL ethyl alcohol and 2mL, mass fraction
36% concentrated hydrochloric acid is extracted, and obtained by drying after centrifuge washing to arrive mesoporous organosilicon hollow nanosphere carrier, grinding is stand-by；
(2) fixed uricase: the mesoporous organosilicon hollow nanosphere carrier for taking 10mg step (1) to grind, 10mL concentration, which is added, is
The uric acid enzyme solution of 0.2mg/mL stirs 8h under 4 DEG C of environment, and taking precipitate freeze-drying is after centrifugation to get arriving carrier-uricase；
(3) glycine for the carrier-uricase for the quality 3mg that 7 parts of steps (2) obtain being added separately to 2mL, pH=8.5 is slow
In the mixed solution of fliud flushing and 0.5mL ultrapure water, then be separately added into 0.5mL concentration be respectively 0.001mg/mL, 0.01mg/mL,
The uric acid titer of 0.05mg/mL, 0.1mg/mL, 0.5mg/mL, 1.0mg/mL, 1.5mg/mL, 40 DEG C of water-bath 20min,
Then absorbance value of the mixed liquor at 290nm is detected respectively using ultraviolet specrophotometer；Concentration with uric acid titer is
Abscissa makes standard curve by ordinate of the absorbance value at the 290nm measured；
(4) carrier-uricase that 3mg step (2) obtains is added to the glycine buffer of 2mL, pH=8.5 and 0.5mL surpasses
In the mixed solution of pure water, the uric acid solution of 0.5mL unknown concentration, 40 DEG C of water-bath 20min, then using ultraviolet are added
Spectrophotometer detects absorbance value of the mixed liquor at 290nm；The standard curve that absorbance value substitution step (3) is obtained
In, the concentration of the uric acid solution is calculated, to realize the detection to uric acid.
2. a kind of method of the fixed uricase detection uric acid of mesoporous organosilicon hollow nanospheres as described in claim 1, special
Sign is: preparing monodisperse SiO using stober method23mL ethyl orthosilicate is added to 37mL ethyl alcohol by nanosphere, 5mL surpasses
In the mixture of pure water and 1.6mL ammonium hydroxide, cleans and dry after stirring centrifugation, obtain monodisperse SiO2Nanosphere.
3. a kind of method of the fixed uricase detection uric acid of mesoporous organosilicon hollow nanospheres as described in claim 1, special
Sign is: the mass fraction of ammonium hydroxide is 28% in step (1).
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111575341A (en) * | 2020-06-29 | 2020-08-25 | 清华大学深圳国际研究生院 | Method for detecting uric acid by using mercaptopropyl agarose beads loaded with uricase and catalase |
| CN111595800A (en) * | 2020-06-08 | 2020-08-28 | 吉林大学 | Immobilized glucose oxidase with aminated dendritic mesoporous silica as carrier and application of immobilized glucose oxidase in detection of glucose |
| CN112924508A (en) * | 2021-01-28 | 2021-06-08 | 合肥工业大学 | Uricase protein inorganic hybrid nanoflower material, electrode, electrochemical sensor, preparation method and application in uric acid monitoring |
| CN113804638A (en) * | 2021-09-17 | 2021-12-17 | 江苏恰瑞生物科技有限公司 | Method for immobilizing activity of uricase |
| CN115254009A (en) * | 2022-09-02 | 2022-11-01 | 江苏恰瑞生物科技有限公司 | Preparation method of blood perfusion device filler for reducing uric acid concentration |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008153157A1 (en) * | 2007-06-15 | 2008-12-18 | Funai Electric Advanced Applied Technology Research Institute Inc. | Enzyme electrode and enzyme sensor |
| CN107019802A (en) * | 2017-03-22 | 2017-08-08 | 南京邮电大学 | A kind of flexible hollow mesoporous organic silicon oxide nanometer capsule material and preparation method |
| CN107753464A (en) * | 2016-08-19 | 2018-03-06 | 牟中原 | It is encapsulated hollow silicon dioxide nano-particle, its preparation method and the application of bioactive ingredients |
| CN109395079A (en) * | 2018-10-17 | 2019-03-01 | 中国人民解放军南京军区南京总医院 | A kind of multifunctional nano probe and its preparation method and application |
-
2019
- 2019-05-31 CN CN201910467336.1A patent/CN110108656A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008153157A1 (en) * | 2007-06-15 | 2008-12-18 | Funai Electric Advanced Applied Technology Research Institute Inc. | Enzyme electrode and enzyme sensor |
| CN107753464A (en) * | 2016-08-19 | 2018-03-06 | 牟中原 | It is encapsulated hollow silicon dioxide nano-particle, its preparation method and the application of bioactive ingredients |
| CN107019802A (en) * | 2017-03-22 | 2017-08-08 | 南京邮电大学 | A kind of flexible hollow mesoporous organic silicon oxide nanometer capsule material and preparation method |
| CN109395079A (en) * | 2018-10-17 | 2019-03-01 | 中国人民解放军南京军区南京总医院 | A kind of multifunctional nano probe and its preparation method and application |
Non-Patent Citations (3)
| Title |
|---|
| HOUBING ZOU等: "An organosilane-directed growth-induced etching strategy for preparing hollow/yolk–shell mesoporous organosilica nanospheres with perpendicular mesochannels and amphiphilic frameworks", 《JOURNAL OF MATERIALS CHEMISTRY A》 * |
| RODOLFO MUNDACA-URIBE等: "Development of a bienzymatic amperometric biosensor to determine uric acid in human serum, based on mesoporous silica (MCM-41) for enzyme immobilization", 《SENSORS AND ACTUATORS》 * |
| 曾艳红: "尿酸、肾上腺素在介孔二氧化硅修饰电极上的电化学行为及测定研究", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111595800A (en) * | 2020-06-08 | 2020-08-28 | 吉林大学 | Immobilized glucose oxidase with aminated dendritic mesoporous silica as carrier and application of immobilized glucose oxidase in detection of glucose |
| CN111575341A (en) * | 2020-06-29 | 2020-08-25 | 清华大学深圳国际研究生院 | Method for detecting uric acid by using mercaptopropyl agarose beads loaded with uricase and catalase |
| CN111575341B (en) * | 2020-06-29 | 2022-07-19 | 清华大学深圳国际研究生院 | Method for detecting uric acid by using mercaptopropyl agarose beads loaded with uricase and catalase |
| CN112924508A (en) * | 2021-01-28 | 2021-06-08 | 合肥工业大学 | Uricase protein inorganic hybrid nanoflower material, electrode, electrochemical sensor, preparation method and application in uric acid monitoring |
| CN112924508B (en) * | 2021-01-28 | 2022-11-18 | 合肥工业大学 | Uricase protein inorganic hybrid nanoflower material, electrode, electrochemical sensor, preparation method and application in uric acid monitoring |
| CN113804638A (en) * | 2021-09-17 | 2021-12-17 | 江苏恰瑞生物科技有限公司 | Method for immobilizing activity of uricase |
| CN115254009A (en) * | 2022-09-02 | 2022-11-01 | 江苏恰瑞生物科技有限公司 | Preparation method of blood perfusion device filler for reducing uric acid concentration |
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