CN105107473A - Preparation method and applications of magnetic nanocomposite particle - Google Patents
Preparation method and applications of magnetic nanocomposite particle Download PDFInfo
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- CN105107473A CN105107473A CN201510435269.7A CN201510435269A CN105107473A CN 105107473 A CN105107473 A CN 105107473A CN 201510435269 A CN201510435269 A CN 201510435269A CN 105107473 A CN105107473 A CN 105107473A
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Abstract
The invention relates to a preparation method and applications of a magnetic nanocomposite particle, and belongs to the technical field of nano material preparation and bioactive substance separation and purification. According to the preparation method, ultrasonic-assisted chemical coprecipitation is adopted to prepare the magnetic nanocomposite particle, wherein Fe3O4 is taken as the core, and chitosan, carboxymethyl chitosan, sodium chondroitin sulfate, and citric acid are taken as shell materials. The magnetic nanocomposite particle possesses abundant surface functional groups, superparamagnetism, and excellent water dispersibility, and is suitable for application in the fields of food and medicine industry; lysozyme purity and activity preservation of a lysozyme purification method based on the magnetic nanocomposite particle are not poorer than that of conventional methods, and the lysozyme purification method based on the magnetic nanocomposite particle is simple in technology, short in production period, low in production cost, and is suitable for industrialized continuous production.
Description
Technical field
The present invention relates to preparation and the bioactivator separating and purifying technology field of nano material, particularly one is with Fe
3o
4for core, the preparation of a kind of magnetic Nano composite granules being shell with native biopolymer polysaccharide or citric acid and the method for one-step method fast separating and purifying lysozyme from egg white thereof.
Background technology
Lysozyme (Lysozyme, EC3.2.1.17) is a kind of hydrolase acting on microorganism wall specially, has the effect of sterilization and broken wall, is a kind of effective antiseptic.Lysozyme range of application is comparatively extensive, all plays an important role in fields such as food, medicine, biology and feed industries.In recent years, market at home and abroad, lysozyme is a kind of very well sold and in short supply bioactivator.
So far, the commercial of practical application is mostly egg white lysozyme, and it is also that structure effect understands the most clearly one of lysozyme, and therefore egg white is the primary raw material of separation and purification lysozyme.At present, the isolation and purification method of the lysozyme reported mainly contains salting out method, crystallisation, ultrafiltration, resin adsorption method etc.As (see CN101768577A) such as Sun Weijun from birds, beasts and eggs, extract lysozyme time, adopt Enzymatic Extraction, separation, freezing, centrifugal, ultrafiltration to obtain lysozyme.When Guo Shuntang etc. (see CN101619308A) extract lysozyme from egg white, then have employed homogeneous, ion-exchange chromatography, the steps such as ultrafiltration obtain lysozyme.Wang Sunan (see CN1844379A) first uses resin adsorption, saline sook afterwards, crystallization have employed egg white during separation and purification lysozyme from egg white, a press filtration, dissolve, secondary press filtration, the steps such as ultrafiltration and three press filtrations just obtain the lysozyme of greater activity.Above purification process is all comparatively complicated, complex operation, easily causes the protein denaturation in egg white at repeatedly process treatment process; In addition, the ultrafiltration production cost used is higher, needs the equipment of special expensive, simultaneously egg white also must with PBS dilute 10 times and more than, make albumen protein be difficult to recycle; Although ion exchange chromatography is applied comparatively extensive at present in the separation and purification of lysozyme, but due to its equipment complexity, complex operation step, post processing easily causes environmental pollution, in leaching process, introduce ammonium sulfate simultaneously, product is not suitable for food pharmaceuticals industry, therefore makes its heavy industrialization apply and is subject to certain restrictions.
Find new lysozyme separating and purifying technology, to reduce production cost, simplify operational sequence, improve purity and the activity of lysozyme product, just become the problem that many researchers are just to be solved.To take magnetic nanoparticle as absorption carrier carry out separation and purification to bioactivator is an emerging separating and purifying technology, surface modified by magnetic nanoparticle, specifically separation and purification target substance, under the effect of externally-applied magnetic field, realizes quick separating from the system of complexity.Compared with conventional method, magnetic Nano isolation technics has simple to operate, quick, the advantage such as efficiently concentrating, high-recovery.
Lysozyme from egg white is made up of 129 amino acid residues, due to wherein containing more alkaline amino acid residue, so its isoelectric point is up to about 10.8, is alkaline enzyme unique in egg white.And there is higher natural activity in neutral salt solution, lysozyme positively charged in neutral conditions under lysozyme normal temperature, therefore can utilize the lysozyme in electrostatic interactions separation and purification egg white.This patent utilizes this peculiar property of lysozyme from egg white exactly, combines the advantage of traditional strong cation-exchanging resin and magnetic Nano isolation technics.
The present invention selects Fe
3o
4for magnetic core, mainly because Fe
3o
4magnetic nanoparticle toxicity is lower, and preparation method is simple, and when particle diameter is less than 30nm, then show superparamagnetism, namely under additional magnetic fields, have stronger magnetic responsiveness, when withdrawing magnetic field, magnetic rapidly disappears, and remanent magnetism is zero, can not be permanently magnetized.The present invention adopts the preparation of the chemical coprecipitation of ultrasonic assistant to synthesize regular appearance, the super-paramagnetism nano composite particles of uniform particle diameter, and the one-step method fast separating and purifying being applied to lysozyme from egg white.
Summary of the invention
The key technical problem that the present invention will solve is the weak point overcoming traditional lysozyme isolation and purification method, a kind of preparation of magnetic Nano composite granules is provided and is applied to the rapid separation and purification method of lysozyme from egg white, the granule-morphology of preparation is regular, uniform particle diameter, magnetic responsiveness are strong, surface functional group enriches, and has good biocompatibility and water dispersible.Lysozyme in magnetic Nano composite granules separation and purification egg white prepared by employing; easy and simple to handle, with low cost; with short production cycle; separative efficiency is high; be separated the lysozyme obtained and there is higher activity; simultaneously technique is easy to amplification and carries out large-scale production lysozyme, solves the problems such as traditional lysozyme separating technology is complicated, step is various.
The preparation method of a kind of magnetic Nano composite granules provided by the invention and application thereof, specific embodiments is as follows:
1, the preparation of magnetic Nano composite granules
Adopt the coprecipitation synthesis of ultrasonic assistant, chemical equation is:
Fe
2++2Fe
3++8NH
3·H
2O=Fe
3O
4↓+8NH
4++4H
2O
A preparation method for magnetic Nano composite granules, carries out according to following step:
By Dissolvable Fe
2+and Fe
3+it is in the mixed solution of 1:1 ~ 10:1 pure water and absolute ethyl alcohol that salt is dissolved in volume ratio, adds surfactant wherein, and logical nitrogen 30min, whole reaction all in ultrasonic system (F=28KHz) carry out; Alkali lye is slowly dripped in reaction system after afterwards temperature being elevated to 40 ~ 70 DEG C, pH value is finally made to reach 10 ~ 11, continue in system, add a certain amount of surface particles material after reaction 30min, afterwards temperature is elevated to 70 ~ 90 DEG C of reaction 1 ~ 6h, cessation reaction, afterwards separating magnetic particles under the effect of externally-applied magnetic field, and by washed with de-ionized water for several times to neutral, and carry out drying process, magnetic Nano composite granules powder, be abbreviated as MNPs.
The Fe of wherein said solubility
2+compound is four water frerrous chloride or ferrous sulfate heptahydrates; The Fe of solubility
3+compound is Iron trichloride hexahydrate; Wherein Fe
2+with Fe
3+mol ratio be 1:2 ~ 2:1, total iron concentration is 0.03 ~ 0.5M.
Wherein said alkali lye selects any one in ammoniacal liquor or NaOH or its mixture.
Wherein said water and the volume ratio of absolute ethyl alcohol are 1:1 ~ 10:1.
Any one in Macrogol 6000, neopelex and softex kw selected by wherein said surfactant.
Wherein said surface particles material is any one in shitosan, CMC, sodium chondroitin sulfate or citric acid.
MNPs regular appearance prepared by said method, uniform particle diameter, surface functional group enriches, and has superparamagnetism, has good dispersiveness and biocompatibility in aqueous simultaneously, can be used for the separation and purification of lysozyme from egg white.Magnetic Nano composite granules is separated under the effect of externally-applied magnetic field, the chemical constituent of not adsorbed and reacting completely not free in the magnetic Nano composite granules of therefore last gained.
, magnetic Nano composite granules is applied to the fast separating and purifying of lysozyme from egg white
(1)
the preparation of fresh albumen stoste
Yolk, egg white are separated, afterwards by fresh albumen and PBS (20mM, pH7.0 ~ 10.0) by volume the ratio of 1:1 ~ 1:3 dilute, then continuous stirring reaction 6h in ice-water bath, stir produce white flock precipitate at 4 DEG C with 10,000rpm refrigerated centrifuge 30min, removing infusible precipitate; Gained supernatant is as the egg white stoste of the separation and purification of lysozyme.
(2)
adopt magnetic Nano composite granules separation and purification lysozyme from egg white stoste
MNPs is joined in egg white stoste, under 0 ~ 37 DEG C of condition, stir (100 ~ 400rpm) and react 30 ~ 180min, the lysozyme in egg white is made specifically to be adsorbed on MNPs surface, under the effect of externally-applied magnetic field, the MNPs being adsorbed with lysozyme is separated from egg white solution, remove impurity three times by washed with de-ionized water, then carry out wash-out with eluent, collect the eluent of lysozyme, eluent is kept in 4 DEG C of refrigerators.Lysozyme eluent adopts SDS-PAGE electrophoresis to carry out purity analysis.Resolving gel concentration: 12%; Concentrated gum concentration: 6%.Finally lysozyme eluent is carried out freeze drying, and measure its active reservation.
Wherein said PBS (pH7.0 ~ 10.0) dilutes fresh albumen.
Wherein said magnetic Nano composite granules and the reaction time of egg white solution are 30 ~ 180min.
PBS (pH=9,20mM, 0.5M sodium chloride) selected by wherein said eluent, PBS (pH=9,20mM, 1M sodium chloride), any one in acetic acid (0.1M) and hydrochloric acid (0.1M).
Wherein said adsorption reaction temperature is 0 ~ 37 DEG C.
Wherein said churned mechanically rotating speed is within the scope of 100 ~ 400rpm.
Antalzyme activity unit definition: under the Lyz optimum reaction conditions of 25 DEG C, the reaction solution system of Lyz and micrococcus lysodeikticus is under 450nm wavelength, and it 0.001 is a unit of activity that one minute internal absorbance often declines.
Beneficial effect of the present invention: the invention provides a kind of preparation of magnetic Nano composite granules and be applied to the rapid separation and purification method of lysozyme from egg white, the magnetic Nano composite granules of preparation is used for the efficient separation and purification of lysozyme from egg white, substantially increase extraction efficiency, farthest maintain the activity of lysozyme simultaneously.The purification process being based upon the lysozyme from egg white on this magnetic Nano composite granules basis is the purity of lysozyme or actively retains all no less than conventional method.
The preparation of a kind of magnetic Nano composite granules provided by the invention and the rapid separation and purification method being applied to lysozyme from egg white thereof have the following advantages:
(1) simple to operate, the separation and purification cycle is shorter:this separation purifying technique adopts magnetic Nano composite granules separation and purification lysozyme from egg white solution, from absorption, cleans to the whole process of wash-out and completes in 2 ~ 3h.
(2) specific adsorption of lysozyme is good, is beneficial to the preparation of high-purity lysozyme:under the purification condition of setting (specific pH value and salinity), magnetic Nano composite granules is by the lysozyme in electrostatic interaction energy specifically separation and purification egg white.
(3) reaction condition is gentle, and high enzyme is lived and retained:this separation purifying technique carries out under the aqueous solution and room temperature condition, and lysozyme remains higher enzyme and lives, and also can not cause the sex change of other protein components in egg white simultaneously.
Detailed description of the invention
Below by embodiment, technical scheme of the present invention is further described:
embodiment 1
0.01M ferrous sulfate heptahydrate and 0.02M Iron trichloride hexahydrate are dissolved in (V/V=10:1) in water-ethanol mixed solution, add Macrogol 6000, logical nitrogen 30min under ultrasonic wave (F=28KHz) dispersity; Temperature is elevated to 40 DEG C, slowly drip ammoniacal liquor afterwards in system, the pH value of regulation system is to about 10, what obtain black contains magnetic particle solution, adds rapidly 2% sodium chondroitin sulfate solution after reaction 30min, is warmed up to 70 DEG C of reaction 1h, cessation reaction, leaves standstill cooling, obtains reactant liquor; Separating magnetic particles under the effect of externally-applied magnetic field, and by washed with de-ionized water for several times to neutral, and carry out freeze drying process, the ferroferric oxide magnetic nano composite particle powder modified of sodium chondroitin sulfate.
Yolk, egg white are separated, afterwards by fresh albumen and PBS (20mM, pH7.0) ratio being 1:1 in V/V is diluted, then continuous stirring reaction 6h in ice-water bath, stir produce white flock precipitate at 4 DEG C with 10,000rpm refrigerated centrifuge 30min, removing infusible precipitate.Gained supernatant is as the egg white stoste of the separation and purification of lysozyme.
The ferroferric oxide magnetic nano composite particles that sodium chondroitin sulfate is modified is joined in egg white stoste, 30min is reacted in the ice-water bath of 0 DEG C, speed of agitator 100rpm/min, the lysozyme in egg white is made specifically to be adsorbed on magnetic nanoparticle surface, under the effect of externally-applied magnetic field, the magnetic nanoparticle being adsorbed with lysozyme is separated from egg white solution, impurity is removed three times by washed with de-ionized water, acetic acid (0.1M) is used to carry out wash-out again, collect the eluent of lysozyme, SDS-PAGE electrophoresis is adopted to carry out purity analysis, resolving gel concentration: 12%; Concentrated gum concentration: 6%.Carry out freeze drying afterwards, measure enzyme and live, relative enzyme lives 86.7%).
embodiment 2
0.2M tetra-water frerrous chloride and 0.1M Iron trichloride hexahydrate are dissolved in (V/V=1:1) in water-ethanol mixed solution, add neopelex, logical nitrogen 30min under ultrasonic wave (F=28KHz) dispersity; Temperature is elevated to 60 DEG C, slowly drip sodium hydroxide solution afterwards in system, the pH value of regulation system is to about 11, what obtain black contains magnetic particle solution, adds rapidly 2% citric acid solution after reaction 30min, is warmed up to 80 DEG C of reaction 1h, cessation reaction, leaves standstill cooling, obtains reactant liquor; Separating magnetic particles under the effect of externally-applied magnetic field, and by washed with de-ionized water for several times to neutral, and carry out freeze drying process, the ferroferric oxide magnetic nano composite particle powder of citric acid modification.
Yolk, egg white are separated, afterwards by fresh albumen and PBS (20mM, pH9.0) ratio being 1:2 in V/V is diluted, then continuous stirring reaction 6h in ice-water bath, stir produce white flock precipitate at 4 DEG C with 10,000rpm refrigerated centrifuge 30min, removing infusible precipitate.Gained supernatant is as the egg white stoste of the separation and purification of lysozyme.
The ferroferric oxide magnetic nano composite particles of citric acid modification is joined in egg white stoste, at 25 DEG C of thermotonus 60min, speed of agitator 200rpm/min, the lysozyme in egg white is made specifically to be adsorbed on magnetic nanoparticle surface, under the effect of externally-applied magnetic field, the magnetic nanoparticle being adsorbed with lysozyme is separated from egg white solution, impurity is removed three times by washed with de-ionized water, use PBS (pH=9 again, 20mM, 0.5M sodium chloride) carry out wash-out, collect the eluent of lysozyme, SDS-PAGE electrophoresis is adopted to carry out purity analysis, resolving gel concentration: 12%, concentrated gum concentration: 6%.Carry out dialysis desalting afterwards, freeze drying process, measure enzyme and live, relative enzyme lives 82.4%).
embodiment 3
0.2M ferrous sulfate heptahydrate and 0.3M Iron trichloride hexahydrate are dissolved in (V/V=5:1) in water-ethanol mixed solution, add Macrogol 6000, logical nitrogen 30min under ultrasonic wave (F=28KHz) dispersity; Temperature is elevated to 70 DEG C, slowly drip sodium hydroxide solution afterwards in system, the pH value of regulation system is to about 11, what obtain black contains magnetic particle solution, add rapidly 1% carboxymethyl chitosan solution after reaction 30min, be warmed up to 90 DEG C of reaction 6h, cessation reaction, leave standstill cooling, obtain reactant liquor; Separating magnetic particles under the effect of externally-applied magnetic field, and by washed with de-ionized water for several times to neutral, and carry out freeze drying process, the sugar-modified ferroferric oxide magnetic nano composite particle powder of carboxymethyl chitosan.
Yolk, egg white are separated, afterwards by fresh albumen and PBS (20mM, pH10.0) ratio being 1:3 in V/V is diluted, then continuous stirring reaction 6h in ice-water bath, stir produce white flock precipitate at 4 DEG C with 10,000rpm refrigerated centrifuge 30min, removing infusible precipitate.Gained supernatant is as the egg white stoste of the separation and purification of lysozyme.
Ferroferric oxide magnetic nano composite particles sugar-modified for carboxymethyl chitosan is joined in egg white stoste, at 37 DEG C of thermotonus 180min, speed of agitator 400rpm/min, the lysozyme in egg white is made specifically to be adsorbed on magnetic nanoparticle surface, under the effect of externally-applied magnetic field, the magnetic nanoparticle being adsorbed with lysozyme is separated from egg white solution, impurity is removed three times by washed with de-ionized water, hydrochloric acid (0.1M) is used to carry out wash-out again, collect the eluent of lysozyme, SDS-PAGE electrophoresis is adopted to carry out purity analysis, resolving gel concentration: 12%; Concentrated gum concentration: 6%.Carry out freeze drying afterwards, measure enzyme and live, relative enzyme lives 78.5%).
Claims (8)
1. a preparation method for magnetic Nano composite granules, is characterized in that carrying out according to following step:
By Dissolvable Fe
2+and Fe
3+it is in the mixed solution of 1:1 ~ 10:1 pure water and absolute ethyl alcohol that salt is dissolved in volume ratio, adds surfactant wherein, and logical nitrogen 30min, whole reaction all in ultrasonic system (F=28KHz) carry out; Alkali lye is slowly dripped in reaction system after afterwards temperature being elevated to 40 ~ 70 DEG C, pH value is finally made to reach 10 ~ 11, continue in system, add a certain amount of surface particles material after reaction 30min, afterwards temperature is elevated to 70 ~ 90 DEG C of reaction 1 ~ 6h, cessation reaction, afterwards separating magnetic particles under the effect of externally-applied magnetic field, and by washed with de-ionized water for several times to neutral, and carry out drying process, magnetic Nano composite granules powder, be abbreviated as MNPs.
2. the preparation method of a kind of magnetic Nano composite granules according to claim 1, is characterized in that the Fe of wherein said solubility
2+compound is four water frerrous chloride or ferrous sulfate heptahydrates; The Fe of solubility
3+compound is Iron trichloride hexahydrate; Wherein Fe
2+with Fe
3+mol ratio be 1:2 ~ 2:1, total iron concentration is 0.03 ~ 0.5M.
3. the preparation method of a kind of magnetic Nano composite granules according to claim 1, is characterized in that wherein said alkali lye selects any one in ammoniacal liquor or NaOH or its mixture.
4. the preparation method of a kind of magnetic Nano composite granules according to claim 1, is characterized in that the volume ratio of wherein said water and absolute ethyl alcohol is 1:1 ~ 10:1.
5. the preparation method of a kind of magnetic Nano composite granules according to claim 1, is characterized in that any one in Macrogol 6000, neopelex and softex kw selected by wherein said surfactant.
6. the preparation method of a kind of magnetic Nano composite granules according to claim 1, is characterized in that wherein said surface particles material is any one in shitosan, CMC, sodium chondroitin sulfate or citric acid.
7. a kind of magnetic Nano composite granules according to claim 1 is applied to the method for the fast separating and purifying of lysozyme from egg white, it is characterized in that carrying out according to following step:
(1)
the preparation of fresh albumen stoste
Yolk, egg white are separated, afterwards by fresh albumen and PBS (20mM, pH7.0 ~ 10.0) by volume the ratio of 1:1 ~ 1:3 dilute, then continuous stirring reaction 6h in ice-water bath, stir produce white flock precipitate at 4 DEG C with 10,000rpm refrigerated centrifuge 30min, removing infusible precipitate; Gained supernatant is as the egg white stoste of the separation and purification of lysozyme;
(2)
adopt magnetic Nano composite granules separation and purification lysozyme from egg white stoste
MNPs is joined in egg white stoste, under 0 ~ 37 DEG C of condition, stir (100 ~ 400rpm) and react 30 ~ 180min, the lysozyme in egg white is made specifically to be adsorbed on MNPs surface, under the effect of externally-applied magnetic field, the MNPs being adsorbed with lysozyme is separated from egg white solution, remove impurity three times by washed with de-ionized water, then carry out wash-out with eluent, collect the eluent of lysozyme, eluent is kept in 4 DEG C of refrigerators;
Lysozyme eluent adopts SDS-PAGE electrophoresis to carry out purity analysis;
Resolving gel concentration: 12%; Concentrated gum concentration: 6%;
Finally lysozyme eluent is carried out freeze drying, and measure its active reservation.
8. a kind of magnetic Nano composite granules according to claim 7 is applied to the method for the fast separating and purifying of lysozyme from egg white, it is characterized in that wherein said PBS (pH7.0 ~ 10.0) dilutes fresh albumen;
Wherein said magnetic Nano composite granules and the reaction time of egg white solution are 30 ~ 180min;
PBS (pH=9,20mM, 0.5M sodium chloride) selected by wherein said eluent, PBS (pH=9,20mM, 1M sodium chloride), any one in acetic acid (0.1M) and hydrochloric acid (0.1M);
Wherein said adsorption reaction temperature is 0 ~ 37 DEG C;
Wherein said churned mechanically rotating speed is within the scope of 100 ~ 400rpm.
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Cited By (9)
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| CN105505900A (en) * | 2016-02-01 | 2016-04-20 | 广西民族大学 | Extraction method of thiol protease |
| CN107132346A (en) * | 2017-05-23 | 2017-09-05 | 天津农学院 | A kind of ferroso-ferric oxide immune nanometer magnetic bead and its preparation method and application |
| CN108499535A (en) * | 2018-04-08 | 2018-09-07 | 长沙理工大学 | A kind of preparation of beta-cyclodextrin-ferriferrous oxide nano magnetic bead |
| CN109651811A (en) * | 2018-11-26 | 2019-04-19 | 宋文星 | Magnetic SF/PEI nano particle and application thereof |
| CN111449957A (en) * | 2020-05-29 | 2020-07-28 | 浙江尖峰健康科技有限公司 | Preparation method of traditional Chinese medicine decoction pieces |
| CN111926003A (en) * | 2020-07-16 | 2020-11-13 | 郑大锋 | Preparation of magnetic microsphere and immobilization of horseradish peroxidase by magnetic microsphere |
| CN112730839A (en) * | 2021-01-20 | 2021-04-30 | 宁波海壹生物科技有限公司 | Kit for measuring content of cytokeratin 19 fragments by magnetic particle chemiluminescence method |
| CN114939187A (en) * | 2022-03-24 | 2022-08-26 | 上海大学 | 3D prints MnPSe 3 Nano-sheet composite support and preparation method and application thereof |
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|---|---|---|---|---|
| CN105505900A (en) * | 2016-02-01 | 2016-04-20 | 广西民族大学 | Extraction method of thiol protease |
| CN107132346A (en) * | 2017-05-23 | 2017-09-05 | 天津农学院 | A kind of ferroso-ferric oxide immune nanometer magnetic bead and its preparation method and application |
| CN108499535A (en) * | 2018-04-08 | 2018-09-07 | 长沙理工大学 | A kind of preparation of beta-cyclodextrin-ferriferrous oxide nano magnetic bead |
| CN109651811A (en) * | 2018-11-26 | 2019-04-19 | 宋文星 | Magnetic SF/PEI nano particle and application thereof |
| CN111449957A (en) * | 2020-05-29 | 2020-07-28 | 浙江尖峰健康科技有限公司 | Preparation method of traditional Chinese medicine decoction pieces |
| CN111449957B (en) * | 2020-05-29 | 2022-04-08 | 浙江尖峰健康科技有限公司 | Preparation method of traditional Chinese medicine decoction pieces |
| CN111926003A (en) * | 2020-07-16 | 2020-11-13 | 郑大锋 | Preparation of magnetic microsphere and immobilization of horseradish peroxidase by magnetic microsphere |
| CN112730839A (en) * | 2021-01-20 | 2021-04-30 | 宁波海壹生物科技有限公司 | Kit for measuring content of cytokeratin 19 fragments by magnetic particle chemiluminescence method |
| CN112730839B (en) * | 2021-01-20 | 2024-01-30 | 宁波海尔施智造有限公司 | Kit for measuring content of cytokeratin 19 fragments by magnetic particle chemiluminescence method |
| CN114939187A (en) * | 2022-03-24 | 2022-08-26 | 上海大学 | 3D prints MnPSe 3 Nano-sheet composite support and preparation method and application thereof |
| CN116035973A (en) * | 2023-01-09 | 2023-05-02 | 杭州大贝生物科技有限公司 | A composition containing flos Camelliae Japonicae extract for relieving allergy and its application in infants Tong Hufu |
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