CN103394338A - Preparation of novel magnetic nano-particle and method for separation of lysozyme from egg white - Google Patents
Preparation of novel magnetic nano-particle and method for separation of lysozyme from egg white Download PDFInfo
- Publication number
- CN103394338A CN103394338A CN2013102535092A CN201310253509A CN103394338A CN 103394338 A CN103394338 A CN 103394338A CN 2013102535092 A CN2013102535092 A CN 2013102535092A CN 201310253509 A CN201310253509 A CN 201310253509A CN 103394338 A CN103394338 A CN 103394338A
- Authority
- CN
- China
- Prior art keywords
- lysozyme
- egg white
- magnetic nanoparticle
- preparation
- sugar
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Peptides Or Proteins (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
Abstract
Belonging to the fields of nano-materials and bioactive substance purification technologies, the invention relates to preparation of a novel magnetic nano-particle and a method for separation of lysozyme from egg white. The invention provides a method for preparation of a carboxymethyl chitosan modified ferroferric oxide magnetic nano-particle by a chemical coprecipitation process, and the magnetic particle is taken as an adsorption carrier for separation and purification of lysozyme in egg white. The nano-magnetic particle prepared by the invention has superparamagnetism and good water dispersibility, and is applicable to food, medical industry and other fields. The lysozyme separation and purification method taking the magnetic nano-particle as the adsorption carrier can achieve a separation effect equal to that of traditional methods.
Description
Technical field
The present invention relates to nano material and bioactivator separating and purifying technology field, particularly a kind of employing have biocompatibility the sugar-modified ferroferric oxide magnetic nanoparticle of natural polysaccharide-carboxymethyl chitosan preparation and from the method for separation and purification lysozyme egg white.
Background technology
Lysozyme (Lysozyme, EC3.2.1.17) is a kind of hydrolase that acts on specially microorganism wall, extensively is present in animal, plant and microbial body, has the effect of sterilization and broken wall.The lysozyme range of application is comparatively extensive, the sterilization of food, fresh-keeping, anticorrosion, antibiotic, antiviral, promote blood clotting, the thalline of fermentation industry to break and the clinical treatment aspect such as tissue repair has also obtained extensive use.At present, market at home and abroad, lysozyme is a kind of very well sold and in short supply biochemical substances.Along with developing rapidly of food, microorganism and biotechnology, and lysozyme is the applying of field of medicaments, and people grow with each passing day to the demand of lysozyme, and are simultaneously, also more and more higher to the requirement of lysozyme quality.The preparation of existing lysozyme is raw material mainly with egg white, and isolation and purification method mainly comprises crystallisation, salting out method, ethanol precipitation, ultrafiltration concentration and ion exchange chromatography, and wherein ion exchange chromatography application at present is comparatively extensive.Yet due to ion-exchange equipment complexity, complex operation, post processing easily causes the existence of the problem such as environmental pollution, and its heavy industrialization application is subject to certain restrictions.Therefore, find new lysozyme separating and purifying technology, to improve purity and the activity of lysozyme product, become many researchers problem just to be solved.Take magnetic nanoparticle as absorption carrier, bioactivator being carried out to separation and purification is an emerging separating and purifying technology, surface modified by to magnetic nanoparticle, specifically the separation and purification target substance, under the effect of externally-applied magnetic field, separate fast from realization the system of complexity.With conventional method, compare the advantage such as that magnetic separation technique has is simple to operate, quick, efficiently concentrating, high-recovery.
Ferroferric oxide magnetic nanoparticle toxicity is lower, and the preparation method is simple, when particle diameter during less than 30nm, show superparamagnetism, namely under additional magnetic fields, have stronger magnetic responsiveness, when withdrawing magnetic field, magnetic rapidly disappears, and remanent magnetism is zero, can be by permanent magnetization.
CMC is a kind of natural polysaccharide with biocompatibility, ferroferric oxide magnetic nanoparticle through the CMC coating, both can make it have good magnetic responsiveness and water dispersible, simultaneously because amino and carboxyl isoreactivity group are contained in its surface, can be further and the various biomolecules couplings such as enzyme, protein, antibody.Under the effect of externally-applied magnetic field, magnetic nanoparticle can separate with reactant liquor easily, has easy and simple to handle and separative efficiency advantages of higher.
Up to the present, the magnetic of employing CMC nano particle is not yet arranged as the separation and purification of absorption carrier for lysozyme from egg white.The present invention adopts the synthetic pattern of chemical coprecipitation preparation regular, the superparamagnetism CMC ferroferric oxide magnetic nanoparticle of uniform particle diameter, and be applied to the separation and purification of lysozyme from egg white.
Summary of the invention
The object of the present invention is to provide the preparation method of the sugar-modified ferroferric oxide magnetic nanoparticle of a kind of carboxymethyl chitosan, the granule-morphology of preparation is regular, uniform particle diameter, magnetic responsiveness are strong, has good biocompatibility.Adopt the lysozyme in prepared magnetic nanoparticle separation and purification egg white, easy and simple to handle, with low cost, with short production cycle, and be easy to the large-scale production lysozyme, solved the problems such as traditional lysozyme separating technology is complicated, step is various.
The method of the preparation of the ferroferric oxide magnetic nanoparticle that carboxymethyl chitosan provided by the invention is sugar-modified and separation and purification lysozyme from egg white thereof, embodiment is as follows:
1. chemical coprecipitation prepares the sugar-modified ferroferric oxide magnetic nanoparticle of carboxymethyl chitosan, and step is as follows:
(1) Fe
2+And Fe
3+Be dissolved in water-absolute ethyl alcohol mixed solution, add surfactant, logical nitrogen 20min, wherein Fe under mechanical agitation
2+Be four water frerrous chloride or ferrous sulfate heptahydrates, Fe
3+For Iron trichloride hexahydrate, Fe
2+And Fe
3+Mol ratio be 1: 2～2: 1, the total concentration of iron ion is 0.01～0.5M, the volume ratio of water and absolute ethyl alcohol is 1: 1～10: 1, surfactant is Macrogol 6000, any one in trisodium citrate and softex kw.
(2) temperature is elevated to 60 ℃, under stirring, slowly drip alkali lye in system, the pH value of regulation system is to 10-11, under stirring, adding rapidly concentration is 1～10% carboxymethyl chitosan sugar juice, be warmed up to 80 ℃ of reaction 1h, cessation reaction, standing cooling, separating magnetic particles under the effect of externally-applied magnetic field, and extremely neutral for several times with washed with de-ionized water, the magnetic-particle of collecting is carried out to drying and process, get final product to obtain the sugar-modified ferroferric oxide magnetic nanoparticle powder of carboxymethyl chitosan.Wherein alkali lye is ammoniacal liquor or NaOH or both mixtures, and stir speed (S.S.) is 600～800rpm.
2. apply the magnetic nanoparticle that makes lysozyme from egg white is separated, its technique is:
(1) with PBS (20mM, pH7.0～10.0) with fresh albumen, carried out mixed diluting in 1: 1 by volume, 100～400rpm continuous stirring reaction 6h in ice-water bath, 4 ℃ 10,000rpm is centrifugal, and 30min removes infusible precipitate, and the gained supernatant is as the egg white stoste of lysozyme separation and purification;
(2) ferroferric oxide magnetic nanoparticle that carboxymethyl chitosan is sugar-modified joins in egg white stoste, 100～400rpm stirring and adsorbing, 10～120min under 0～37 ℃, absorption is had to the magnetic nanoparticle of lysozyme under the effect of externally-applied magnetic field from egg white solution, separating, with washed with de-ionized water, remove impurity three times, with eluent, carrying out Shen takes off, wherein eluent is selected phosphate buffer (pH=9, 20mM, 0.5M sodium chloride), PBS (pH=9, 20mM, 1M sodium chloride), any in acetic acid (0.1M) and hydrochloric acid (0.1M), collect the eluent of lysozyme, after drying, can obtain lysozyme product.Beneficial effect of the present invention:
(1) the invention provides a kind of method that chemical coprecipitation prepares the sugar-modified ferroferric oxide magnetic nanoparticle of carboxymethyl chitosan, the magnetic nanoparticle pattern that makes is regular, uniform particle diameter, average grain diameter is in the 15nm left and right, has superparamagnetism, simultaneously in the aqueous solution, have good dispersiveness, can be used for the separation and purification of lysozyme from egg white.
(2) separation method of lysozyme from egg white provided by the invention is simple to operate, the separation and purification cycle is shorter, from absorption, clean to the whole process of wash-out and complete in 2～3h.
(3) magnetic nanoparticle take CMC as the finishing thing is by the lysozyme generation specific adsorption in electrostatic interaction energy and egg white, and productive rate and the purity of gained lysozyme product are higher.
(4) lysozyme from egg white separating technology reaction condition gentleness provided by the invention, the antalzyme activity retention rate is higher.
(5) in this technique, the absorption of lysozyme, cleaning and elution process can be carried out the serialization production operation, and whole technique can be carried out the industrialization continued operation.
The accompanying drawing explanation
The transmission electron microscope picture of the ferroferric oxide magnetic nanoparticle that Fig. 1 carboxymethyl chitosan is sugar-modified.
Fourier's infrared spectrogram of the ferroferric oxide magnetic nanoparticle that Fig. 2 carboxymethyl chitosan is sugar-modified.
The SDS-PAGE electrophoretogram of the sugar-modified ferroferric oxide magnetic nanoparticle of Fig. 3 carboxymethyl chitosan to the lysozyme from egg white separation and purification.
The specific embodiment
Embodiment 1
By 0.1MFe
2+And 0.2MFe
3+Be dissolved in (v/v=10: 1), add Macrogol 6000, logical nitrogen 20min under mechanical agitation in the water-ethanol mixed solution.Temperature is elevated to 60 ℃, under stirring, slowly drip ammoniacal liquor in system, the pH value of regulation system is to 10 left and right, the carboxymethyl chitosan sugar juice that adds rapidly 5% concentration under stirring, be warmed up to 80 ℃ of reaction 1h, speed of agitator 600rpm, cessation reaction, standing cooling, separating magnetic particles under the effect of externally-applied magnetic field, and with washed with de-ionized water, for several times to neutral, the magnetic-particle of collecting is carried out to the freeze drying processing, make the sugar-modified ferroferric oxide magnetic nanoparticle powder of carboxymethyl chitosan.
Fresh albumen and PBS (20mM, pH7.0) are diluted in the ratio of 1: 1, then continuous stirring reaction 6h in ice-water bath, 4 ℃ with 10, the centrifugal 30min of 000rpm, remove infusible precipitate, and the gained supernatant is as the egg white stoste source of lysozyme separation and purification.
The ferroferric oxide magnetic nanoparticle that the carboxymethyl chitosan that makes is sugar-modified joins in egg white stoste, at 25 ℃ of thermotonus 120min, speed of agitator 200rpm, absorption is had to the magnetic nanoparticle of lysozyme under the effect of externally-applied magnetic field from egg white solution, separating, with washed with de-ionized water, remove impurity three times, then use PBS (pH=9,20mM, 0.5M sodium chloride) carry out wash-out, collect the eluent of lysozyme, freeze drying obtains lysozyme product.
Embodiment 2
by 0.2MFe
2+and 0.1MFe
3+be dissolved in (v/v=5: 1) in the water-ethanol mixed solution, add trisodium citrate, logical nitrogen 20min under mechanical agitation, temperature is elevated to 60 ℃, under stirring, slowly drip sodium hydroxide solution in system, the pH value of regulation system is to 10 left and right, the carboxymethyl chitosan sugar juice that adds rapidly 2% concentration under stirring, be warmed up to 80 ℃ of reaction 1h, speed of agitator 800rpm, cessation reaction, standing cooling, separating magnetic particles under the effect of externally-applied magnetic field, and extremely neutral for several times with washed with de-ionized water, the magnetic-particle of collecting is carried out to the freeze drying processing, make the sugar-modified ferroferric oxide magnetic nanoparticle powder of carboxymethyl chitosan.
Fresh albumen and PBS (20mM, pH9.0) are diluted in the ratio of 1: 1, continuous stirring reaction 6h in ice-water bath then, 4 ℃ with the centrifugal 30min of 10,000rpm, removes infusible precipitate.The gained supernatant is as the egg white stoste source of lysozyme separation and purification.
The ferroferric oxide magnetic nanoparticle that the carboxymethyl chitosan that makes is sugar-modified joins in egg white stoste, at 37 ℃ of thermotonus 60min, speed of agitator 200rpm, absorption is had to the magnetic nanoparticle of lysozyme under the effect of externally-applied magnetic field from egg white solution, separating, with washed with de-ionized water, remove impurity three times, use again acetic acid (0.1M) to carry out wash-out, collect the eluent of lysozyme.Freeze drying obtains lysozyme product.
Claims (7)
1. the preparation of a novel magnetic nano particle and the method for separating egg white lysozyme thereof, it is characterized in that adopting chemical coprecipitation, with the sugar-modified tri-iron tetroxide of carboxymethyl chitosan, make magnetic nanoparticle, and take this particle as absorption carrier, separate the lysozyme in egg white, the preparation technology of magnetic nanoparticle is:
(1) by the Fe of solubility
2+And Fe
3+In the be dissolved in volume ratio water-absolute ethyl alcohol mixed solution of 1: 1～10: 1, add surfactant, logical nitrogen 20min under mechanical agitation;
(2) temperature is elevated to 60 ℃, under stirring, slowly drip alkali lye in system, the pH value to 10 of regulation system～11, under stirring, adding rapidly concentration is 1～10% carboxymethyl chitosan sugar juice, be warmed up to 80 ℃ of reaction 1h, cessation reaction, standing cooling, obtain reactant liquor, separating magnetic particles under the effect of externally-applied magnetic field, and with washed with de-ionized water, for several times to neutral, the magnetic-particle of collecting carried out to drying and process, get final product to obtain the sugar-modified ferroferric oxide magnetic nanoparticle powder of carboxymethyl chitosan;
The technique that the magnetic nanoparticle that application makes separates lysozyme from egg white is:
(1) with PBS (20mM, pH7.0～10.0) with fresh albumen, carried out mixed diluting in 1: 1 by volume, continuous stirring reaction 6h in ice-water bath, 4 ℃ 10,000rpm is centrifugal, and 30min removes infusible precipitate, and the gained supernatant is as the egg white stoste of lysozyme separation and purification;
(2) ferroferric oxide magnetic nanoparticle that carboxymethyl chitosan is sugar-modified joins in egg white stoste, at 0～37 ℃ of lower stirring and adsorbing 10～120min, absorption is had to the magnetic nanoparticle of lysozyme under the effect of externally-applied magnetic field from egg white solution, separating, with washed with de-ionized water, remove impurity three times, with eluent, carry out wash-out, collect the eluent of lysozyme, after drying, can obtain lysozyme product.
2. the preparation method of the sugar-modified ferroferric oxide magnetic nanoparticle of carboxymethyl chitosan according to claim 1, is characterized in that the Fe of solubility
2+Compound is four water frerrous chloride or ferrous sulfate heptahydrates; The Fe of solubility
3+Compound is Iron trichloride hexahydrate; Fe wherein
2+With Fe
3+Mol ratio be 1: 2～2: 1, total iron concentration is 0.01～0.5M.
3. the preparation method of the sugar-modified ferroferric oxide magnetic nanoparticle of carboxymethyl chitosan according to claim 1, is characterized in that alkali lye used is ammoniacal liquor or NaOH or both mixtures.
4. the preparation method of the sugar-modified ferroferric oxide magnetic nanoparticle of carboxymethyl chitosan according to claim 1, is characterized in that surfactant selects Macrogol 6000, any in trisodium citrate and softex kw.
5. the preparation method of the sugar-modified ferroferric oxide magnetic nanoparticle of carboxymethyl chitosan according to claim 1, is characterized in that the speed that stirs described in preparation technology is 600～800rpm.
6. the separating technology of lysozyme from egg white according to claim 1, it is characterized in that eluent selects PBS (pH=9,20mM, 0.5M sodium chloride), PBS (pH=9,20mM, 1M sodium chloride), any in acetic acid (0.1M) and hydrochloric acid (0.1M).
7. the separating technology of lysozyme from egg white according to claim 1, is characterized in that the speed that stirs described in separating technology is 100～400rpm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2013102535092A CN103394338A (en) | 2013-06-20 | 2013-06-20 | Preparation of novel magnetic nano-particle and method for separation of lysozyme from egg white |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2013102535092A CN103394338A (en) | 2013-06-20 | 2013-06-20 | Preparation of novel magnetic nano-particle and method for separation of lysozyme from egg white |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103394338A true CN103394338A (en) | 2013-11-20 |
Family
ID=49558169
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2013102535092A Pending CN103394338A (en) | 2013-06-20 | 2013-06-20 | Preparation of novel magnetic nano-particle and method for separation of lysozyme from egg white |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103394338A (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103990443A (en) * | 2014-03-27 | 2014-08-20 | 华南师范大学 | Single-stranded DNA nucleic acid modified chitosan magnetic microsphere preparation method |
| CN104910445A (en) * | 2015-06-09 | 2015-09-16 | 天津大学 | Chitosan-coated ferroferric oxide magnetic nano composite particle and preparation method thereof |
| CN105056912A (en) * | 2015-07-23 | 2015-11-18 | 江苏大学 | Preparation method for immobilized metal ion affinity magnetic nanoparticles and application thereof |
| CN105107473A (en) * | 2015-07-23 | 2015-12-02 | 江苏大学 | Preparation method and applications of magnetic nanocomposite particle |
| CN105268416A (en) * | 2015-09-30 | 2016-01-27 | 吉林大学 | Preparation method for separating lysozyme by amino-functionalized magnetic nanoparticles |
| CN103990443B (en) * | 2014-03-27 | 2016-11-30 | 华南师范大学 | A kind of single stranded DNA nucleic acid modifies the preparation method of chitosan magnetic microsphere |
| CN107051372A (en) * | 2017-03-28 | 2017-08-18 | 山西清泽阳光环保科技有限公司 | A kind of preparation method of magnetic adsorptive material |
| CN107486156A (en) * | 2017-09-14 | 2017-12-19 | 江南大学 | A kind of method of magnetic nanoparticle adsorbing separation bovine serum albumin(BSA) |
| CN108371942A (en) * | 2018-02-12 | 2018-08-07 | 湖北工业大学 | A kind of composite magnetic nano-particle Fe3O4@Au/MPA/NTA-Ni2+And its it prepares and applies |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6344884A (en) * | 1986-08-11 | 1988-02-25 | Fuji Boseki Kk | Affinity chromatography carrier for purifying lysozyme |
| CN1299869A (en) * | 2001-01-04 | 2001-06-20 | 华南农业大学 | Method of extracting lysozyme from salted egg white |
-
2013
- 2013-06-20 CN CN2013102535092A patent/CN103394338A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6344884A (en) * | 1986-08-11 | 1988-02-25 | Fuji Boseki Kk | Affinity chromatography carrier for purifying lysozyme |
| CN1299869A (en) * | 2001-01-04 | 2001-06-20 | 华南农业大学 | Method of extracting lysozyme from salted egg white |
Non-Patent Citations (1)
| Title |
|---|
| 孙俊: ""功能性磁性纳米颗粒的制备及其在食品体系中的应用研究"", 《中国博士学位论文全文数据库工程科技Ⅰ辑》, no. 07, 15 July 2012 (2012-07-15) * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103990443A (en) * | 2014-03-27 | 2014-08-20 | 华南师范大学 | Single-stranded DNA nucleic acid modified chitosan magnetic microsphere preparation method |
| CN103990443B (en) * | 2014-03-27 | 2016-11-30 | 华南师范大学 | A kind of single stranded DNA nucleic acid modifies the preparation method of chitosan magnetic microsphere |
| CN104910445A (en) * | 2015-06-09 | 2015-09-16 | 天津大学 | Chitosan-coated ferroferric oxide magnetic nano composite particle and preparation method thereof |
| CN105056912A (en) * | 2015-07-23 | 2015-11-18 | 江苏大学 | Preparation method for immobilized metal ion affinity magnetic nanoparticles and application thereof |
| CN105107473A (en) * | 2015-07-23 | 2015-12-02 | 江苏大学 | Preparation method and applications of magnetic nanocomposite particle |
| CN105268416A (en) * | 2015-09-30 | 2016-01-27 | 吉林大学 | Preparation method for separating lysozyme by amino-functionalized magnetic nanoparticles |
| CN107051372A (en) * | 2017-03-28 | 2017-08-18 | 山西清泽阳光环保科技有限公司 | A kind of preparation method of magnetic adsorptive material |
| CN107051372B (en) * | 2017-03-28 | 2019-10-18 | 山西清泽阳光环保科技有限公司 | A kind of preparation method of magnetic adsorptive material |
| CN107486156A (en) * | 2017-09-14 | 2017-12-19 | 江南大学 | A kind of method of magnetic nanoparticle adsorbing separation bovine serum albumin(BSA) |
| CN107486156B (en) * | 2017-09-14 | 2019-10-25 | 江南大学 | A kind of method of magnetic nanoparticle adsorbing separation bovine serum albumin(BSA) |
| CN108371942A (en) * | 2018-02-12 | 2018-08-07 | 湖北工业大学 | A kind of composite magnetic nano-particle Fe3O4@Au/MPA/NTA-Ni2+And its it prepares and applies |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103394338A (en) | Preparation of novel magnetic nano-particle and method for separation of lysozyme from egg white | |
| CN105107473A (en) | Preparation method and applications of magnetic nanocomposite particle | |
| CN100475707C (en) | Method for preparing magnetic Nano composite granules coated by cationic polyelectrolyte | |
| CN105566489B (en) | Method for preparing lactoferrin with different iron saturation degrees | |
| CN104888705B (en) | The preparation method of magnetic iron oxide/bagasse active carbon | |
| CN101343083A (en) | Magnetic carbon sphere of surface finished C8 alkyl chain, preparation and application thereof | |
| CN104480101B (en) | Preparation of magnetic nano particle immobilized amino-acylase and products thereof and application | |
| CN101176506A (en) | Method for promoting dissolve of soy protein | |
| CN103160482A (en) | Method for preparing egg white lysozyme and active protein by adopting coseparation | |
| CN105012940A (en) | Preparation method of nanometer collagen peptide zinc chelate | |
| CN105056912A (en) | Preparation method for immobilized metal ion affinity magnetic nanoparticles and application thereof | |
| CN105724784A (en) | Polypeptide zinc chelate prepared with soy protein isolate as base and preparation method | |
| CN103464126A (en) | Preparation of zirconium dioxide and ferriferrous oxide nanoparticles and method used for enrichment of phosphopeptides by using zirconium dioxide and ferriferrous oxide nanoparticles | |
| CN105661049A (en) | Polypeptide copper chelate prepared by using soybean protein isolate as substrate and preparation method | |
| CN102000342B (en) | Preparation method of superparamagnetism conductive nano gamma-ferric oxide/ polyaniline-dexamethasone sodium phosphate | |
| CN103695513B (en) | A kind of method improving yield of soybean peptide with low molecular weight | |
| CN103464092A (en) | Preparation of novel TiO2/diatomite composite material, and method used for purifying phosvitin phosphopeptides with novel TiO2/diatomite composite material | |
| CN106011206A (en) | Method for preparing bioactive peptide from immobilized bi-enzyme of composite carrier magnetic nano-particles | |
| CN104591410B (en) | A kind of for composite flocculation agent processing low temperature and low turbidity water and preparation method thereof | |
| CN115336757B (en) | Method for preparing water-soluble soybean polysaccharide calcium chelate with assistance of high-voltage pulse electric field | |
| CN113295747B (en) | Application of biosensor chip in lysine detection | |
| CN110506940A (en) | It is a kind of deliver ferrous ion fluorescent nano particles preparation method and its preparation nanometer ferrous ion compound in application | |
| CN105399816A (en) | Method for separating human lactoferrin from transgenic goat milk | |
| CN105018553A (en) | Phosvitin phosphopeptide enrichment method | |
| CN105483201B (en) | Fermentation preparation method of whey antioxidant peptide |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20131120 |