CN108299552A - The extracting method of goat dairy milk fat globule membrane protein for proteomics research - Google Patents
The extracting method of goat dairy milk fat globule membrane protein for proteomics research Download PDFInfo
- Publication number
- CN108299552A CN108299552A CN201810163762.1A CN201810163762A CN108299552A CN 108299552 A CN108299552 A CN 108299552A CN 201810163762 A CN201810163762 A CN 201810163762A CN 108299552 A CN108299552 A CN 108299552A
- Authority
- CN
- China
- Prior art keywords
- membrane protein
- milk fat
- fat globule
- globule membrane
- goat dairy
- 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
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
Abstract
The extracting method of the goat dairy milk fat globule membrane protein for proteomics research of the present invention belongs to the technical field of milk fat globule membrane protein extraction.Sucrose is added into goat dairy sample first, the butter oil of crude separation, the butter oil priority potassium phosphate buffer solution and milli-Q water that will slightly carry are obtained after high speed centrifugation, then are cracked with lysate, it is finally purified with precooling acetone soln, obtains milk fat globule membrane protein sample.The method of the present invention can effectively reduce the influence of low content Caseinum componemt, and the milk fat globule membrane protein recovery rate more high stability extracted is stronger, has preferable popularizing application prospect and economic value in terms of goat dairy milk fat globule membrane protein matter group research.
Description
Technical field
The present invention relates to a kind of using proteome analysis as the extracting method of purpose milk fat globule membrane protein, especially
A kind of extracting method for milk fat globule membrane protein in goat dairy sample.
Background technology
Proteomics research is goed deep into execution body-protein of vital movement in proteomics level
Systematic research, including protein expression level, amino acid sequence, post translational processing and protein interaction, in protein
The various functions of cell, the pathologic process etc. of various physiological and biochemical procedures and disease are understood in level.
Milk fat ball film is the thin film being enclosed in outside butter oil ball, and cross-sectional diameter is about 10~20nm, is risen
The effect of emulsifier, and prevent the polymerization of fat globule and enzyme from degenerating.There are 25%~70% memebrane proteins on milk fat ball film, claim
For milk fat globule membrane protein (Milk fat globule membrane protein.Milk fat globule membrane protein accounts for total protein in breast
The 1%~2% of content, although MFGM albumen content in breast is little, have antitumaous effect, antibiotic property and block resistance with
And to the important biomolecules meaning such as autoimmunization of encephalomyelitis.
In proteomics research, the step of the most critical of the extraction process of sample, the success or failure of experiment are directly determined.By
In the particularity of proteomics research purpose, other goat dairy breasts for the purpose of dairy products manufacture, conventional physical and chemical analysis etc.
The preparation processing step of fat globule membrane proteins is not fully suitable at the preparation of the sample for the purpose of proteomics research
Reason；The milk sample in different genera source is because it is also differed at the different requirement to processing step and parameter being grouped as, mountain at present
One of the main problem that the research of sheep breast milk fat globule membrane protein group faces is exactly that the extraction difficulty of protein group is larger.Mainly
There is following reason：1. milk fat globule membrane protein content is considerably less, wherein (protein is rich for the lower low abundance proteins of content
Degree is height of certain protein in Proteomics content, is usually low in the protein definition of ng/ml~pg/ml by concentration
Abundant protein) it is easy to be covered by other impurities component；2. from " butter oil-milk fat ball film-milk fat globule membrane protein "
Easily occur to extract incomplete situation in extraction process or reduce the stability of protein, impacts proteomics measurement
As a result uncertain factor.
Invention content
The technical problem to be solved by the present invention is to overcome existing using proteomics as research purpose, goat dairy breast
Protein is not easy to extract in fat globule membrane proteins extraction process, the low problem of stability, and it is strong and can have to provide a kind of stability
The sample preparation processing method for the goat dairy milk fat globule membrane protein group research that effect removal impurity composition influences.
To achieve the above object, the technical solution that the present invention provides is as follows.
A kind of extracting method of goat dairy milk fat globule membrane protein for proteomics research, there is following process step
Suddenly；
1) Mining takes goat dairy as whole milk samples；
2) sucrose being added in whole milk samples is stirred, and obtains uniform goat dairy sucrose solution；Sucrose addition
It is the 4% of whole milk samples by mass；
3) goat dairy sucrose solution under the conditions of 4 DEG C is centrifuged into 1h, centrifugal force 50000g, collects upper layer butter oil layer；
4) it uses potassium phosphate buffer solution to be cleaned by ultrasonic butter oil layer, takes upper layer butter oil, it is super to repeat potassium phosphate buffer solution
Sound cleans 1 time；It is cleaned by ultrasonic 1 time with ultra-pure water again, is centrifugally separating to obtain the crude extract of butter oil；Each cleaning phosphoric acid used
Sylvite buffer solution, ultra-pure water are 3 times of upper layer butter oil by volume；
5) crude extract of butter oil is stood to butter oil under the conditions of 4 DEG C and is solidified；
6) the butterfat solids heating water bath of solidification is melted into cream；
7) lysate is added into liquid cream and is stored at room temperature 50~60min, lysate addition is liquid milk oil volume
0.1 times, then centrifugation removal upper layer grease, collect lower sediment, obtain the milk fat globule membrane protein solution of goat dairy；Institute
The lysate stated, composition and mol ratio are dithiothreitol (DTT)s:Tris-HCl:Lauryl sodium sulfate=15:1:60~63, and
A concentration of 1.5mM of the dithiothreitol (DTT) in lysate；
8) pre- cold acetone precipitation：Pre- cold acetone mixing, the temperature of pre- cold acetone are added into milk fat globule membrane protein solution
It it is -20 DEG C, dosage is 5 times of milk fat globule membrane protein liquor capacity；It is placed 10~15 hours at -20 DEG C；Supernatant is abandoned in centrifugation,
Then it is washed 2 times with isometric pre- cold acetone, then sucks supernatant after high speed centrifugation, air-dried precipitation and obtain goat dairy butter oil
Ball memebrane protein.
In step 1), the whole milk samples are the fresh goat dairies that Mining takes；Or at -20 DEG C in 1~3 hour
The goat dairy transported back at~4 DEG C；Or be positioned over to store under -80 DEG C of environment and place, before extraction goat dairy milk fat globule membrane protein
Place it in 4 DEG C of refrigerator defrostings.
In step 6), the heating water bath, bath temperature is 40~50 DEG C.
In step 8), the pre- cold acetone mixing of the addition carries out under ice bath；The air-dried precipitation, wind
The dry time is 5~10min.
Compared with prior art, the beneficial effects of the invention are as follows：First, " potassium phosphate buffer solution-ultra-pure water " is utilized
Goat dairy butter oil layer is washed, the casein and lactalbumin being attached on butter oil layer can be effectively eluted；Secondly, cracking is utilized
Liquid cracks goat dairy cream, can effectively improve the recovery rate of milk fat globule membrane protein, and not to protein itself property
Matter impacts, and can effectively remove influence of the impurity composition (such as low component casein) to milk fat globule membrane protein, make albumen
Property is stablized.In short, the recovery rate of the method milk fat globule membrane protein of the present invention is high, and the milk fat globule membrane protein extracted
Matter is stablized, hence it is evident that improves the purity of sheep breast milk fat globule membrane protein.
Description of the drawings
Fig. 1 is the extraction flow chart of goat dairy milk fat globule membrane protein of the present invention.
Fig. 2 is the figure compared with the albumen concentration for the goat dairy milk fat globule membrane protein that comparative example is extracted of embodiment 1.
Specific implementation mode
With reference to specific embodiment, present invention is further described in detail, to enable those skilled in the art with reference to explanation
Book word can be implemented according to this, but not as a limitation of the invention.
Embodiment 1：A kind of extracting method of goat dairy milk fat globule membrane protein for proteomics research
1) the fresh sheep breast for selecting healthy goat chooses the sheep breast of 30 goats as full milk sample to eliminate individual difference
Product, every goat take 50ml sheep breast, the fresh sheep breast taken from pasture must as early as possible 4 DEG C or -20 DEG C transport laboratory treatment back,
It is such as unable to preserve under the conditions of timely processing need to be positioned over -80 DEG C, 4 DEG C of refrigerator defrostings is placed it in before use, this method must make
With the sheep breast of (- 80 DEG C) preservations of fresh or condition of ultralow temperature, 4 DEG C and -20 DEG C are only applicable to transport in short-term, it is impossible to be used in sheep milk sample
The storage of product is placed, and the degradation of albumen otherwise can occur and influence the measurement result of protein science.
2) it is separately added into the sucrose of 4% (mass fraction) into 30 parts of goat dairy samples in step 1), uses magnetic agitation
Device is stirred, and obtains uniform goat dairy sucrose solution.This step is completed at room temperature, and the operating time should shorten as possible, subtract
Few goat dairy sample is exposed to the time under room temperature environment.
3) the goat dairy sucrose solution obtained in step 2) is centrifuged into 1h, centrifugal force 50000g under the conditions of 4 DEG C, carefully
Collect upper layer butter oil layer.Centrifuging temperature should not increase in this step, and the subsequent operation carried out in room temperature can be led to by increasing temperature
(operation for scraping off fat deposit at ambient temperature needs to consume the regular hour, especially in needs largely processing samples
Wait, this section operation time in can cause centrifugation after milk sample heating) when occur fat deposit back dissolving.
4) it is cleaned by ultrasonic the butter oil layer obtained in step 3) with 3 times of volume potassium phosphate buffer solutions, 4 DEG C after the completion of cleaning
Under the conditions of take upper layer butter oil, which cleans 2 times repeatedly.It is cleaned by ultrasonic 1 with the ultra-pure water of 3 times of volumes again after the completion of cleaning
It is secondary, reaction system is replaced into ultrapure aqueous systems by potassium phosphate buffer system, the crude extract of butter oil is obtained after centrifugation.
5) (about 12 hours) make butter oil solidify overnight under the conditions of the crude extract of butter oil being statically placed in 4 DEG C.
6) the butterfat solids heating water bath of solidification is melted into liquid cream, bath temperature is 40 DEG C.The step temperature
Unsuitable excessively high, temperature can make the stability of lactoprotein reduce higher than 50 DEG C.
7) lysate of 0.1 times of liquid milk oil volume is added into the liquid cream in step 6), is stored at room temperature cracking 50
~60min, then the grease on centrifugation removal upper layer, collects lower sediment, obtains the milk fat globule membrane protein solution of goat dairy.No
Test result can be had an impact with the proportioning of lysate and lysate ingredient, the milk sample in different genera source is because of its component physics and chemistry
The applicable lysate of the difference of property is different, can the lysate that source milk sample of the same race is applicable in by the difference of research purpose
It can be different.The dithiothreitol (DTT) lysate used in this step be obtained after test of many times optimizes be best suited for Goat milk
The lysate of fat globule membrane proteins extraction, the configuration method of the lysate are that will weigh suitable dithiothreitol (DTT) to be dissolved in
Make the final concentration of 1.5mM of dithiothreitol (DTT), the two mixing equal in 0.1mM Tris-HCl (trishydroxymethylaminomethane) buffer solution
40g/L is pressed after even again, lauryl sodium sulfate is added, with magnetic stirrer mixing.
8) pre- cold acetone precipitation：- 20 DEG C of precoolings of 5 times of volumes are added into the milk fat globule membrane protein obtained in step 7)
Acetone, rapid mixing (this process should be carried out in ice bath)；(10~15 hours) are stood overnight at -20 DEG C, high speed centrifugation is abandoned
Clearly, it is then washed twice with isometric pre- cold acetone, supernatant is carefully sucked after high speed centrifugation, uncaps and volatilizes, air-dried precipitation is
Obtaining the goat dairy milk fat globule membrane protein that this method is extracted, (air-dry time is 5~10min, should not be over-drying, if air-dried
Overlong time, the milk fat globule membrane protein of precipitation are likely to occur undissolved situation when redissolving).
It has selected " potassium phosphate buffer solution-ultra-pure water " to wash goat dairy butter oil layer in the present embodiment, has then utilized
Special lysate splits goat dairy cream.Compared with following comparative example, the butter oil of the method extraction of the present embodiment
When ball memebrane protein carries out proteome analysis, pass through a nanoliter tablets by HPLC-MS (nano HPLC-MS/MS)
It detects the protein more than 200 kinds, that is, detects that kinds of protein number is most, the goat dairy milk fat globule membrane protein of extraction
Concentration is about 0.52mg/ml (referring to Fig. 2).
Comparative example 1：Goat dairy cream is not cracked with lysate
1) the fresh sheep breast for selecting healthy goat chooses the sheep breast of 30 goats as full milk sample to eliminate individual difference
Product, every goat take 50ml sheep breast, the fresh sheep breast taken from pasture must as early as possible 4 DEG C or -20 DEG C transport laboratory treatment back,
It is such as unable to preserve under the conditions of timely processing need to be positioned over -80 DEG C, 4 DEG C of refrigerator defrostings is placed it in before use, this method must make
With the sheep breast of (- 80 DEG C) preservations of fresh or condition of ultralow temperature, 4 DEG C and -20 DEG C are only applicable to transport in short-term, it is impossible to be used in sheep milk sample
The storage of product is placed, and the degradation of albumen otherwise can occur and influence the measurement result of proteomics.
2) it is separately added into the sucrose of 4% (mass fraction) into 30 parts of goat dairy samples in step 1), uses magnetic agitation
Device is stirred, and obtains uniform goat dairy sucrose solution.This step is completed at room temperature, and the operating time should shorten as possible, subtract
Few goat dairy sample is exposed to the time under room temperature environment.
3) the goat dairy sucrose solution obtained in step 2) is centrifuged into 1h, centrifugal force 50000g under the conditions of 4 DEG C, carefully
Collect upper layer butter oil layer.Centrifuging temperature should not increase in this method, and the subsequent operation carried out in room temperature can be led to by increasing temperature
(operation for scraping off fat deposit at ambient temperature needs to consume the regular hour, especially in needs largely processing samples
Wait, this section operation time in can cause centrifugation after milk sample heating) when occur fat deposit back dissolving.
4) it is cleaned by ultrasonic the butter oil layer obtained in step (3) with 3 times of volume potassium phosphate buffer solutions, 4 after the completion of cleaning
Upper layer butter oil is taken under the conditions of DEG C, which cleans 2 times repeatedly.It is cleaned by ultrasonic again with the ultra-pure water of 3 times of volumes after the completion of cleaning
Once, reaction system is replaced into ultrapure aqueous systems by potassium phosphate buffer system, the thick of butter oil is obtained after centrifugation and is carried
Object.
5) (about 12 hours) make butter oil solidify overnight under the conditions of the crude extract of butter oil being statically placed in 4 DEG C.
6) the butterfat solids heating water bath of solidification is melted into liquid cream, bath temperature is 40 DEG C.The step temperature
Unsuitable excessively high, temperature can make the stability of lactoprotein reduce higher than 50 DEG C.
7) suitable ultra-pure water is added into the liquid cream in step 6) and is stored at room temperature 50-60min, then centrifugation removal
The grease on upper layer collects lower sediment, obtains the milk fat globule membrane protein of goat dairy.
8) pre- cold acetone precipitation：- 20 DEG C that 5 times of volumes are added in the milk fat globule membrane protein obtained into step (7) are pre-
Cold acetone, rapid mixing (this process should be carried out in ice bath)；(10-15 hours) are stood overnight at -20 DEG C, high speed centrifugation is abandoned
Clearly, it is then washed twice with isometric pre- cold acetone, supernatant is carefully removed after high speed centrifugation, uncaps and volatilizes, air-dried precipitation is
Obtaining the goat dairy milk fat globule membrane protein that this method is extracted, (air-dry time is 5~10min, should not be over-drying, if air-dried
Overlong time, the milk fat globule membrane protein of precipitation are likely to occur undissolved situation when redissolving).
It is measured through BCA protein detection kits (sigma companies), the goat dairy milk fat globule membrane protein concentration of extraction is about
0.26mg/ml (referring to Fig. 2).
Comparative example 2：" potassium phosphate buffer solution-ultra-pure water " useless washs goat dairy butter oil layer
Step 4) is cleaned by ultrasonic the butter oil layer obtained in step 3) with 3 times of volume ultra-pure waters in this comparative example, has cleaned
At upper layer butter oil is taken under the conditions of latter 4 DEG C, which cleans 3 times repeatedly, and the crude extract of butter oil is obtained after centrifugation.Remaining
Step is same as Example 1.It is measured through BCA protein detection kits (sigma companies), the goat dairy milk fat ball film egg of extraction
White concentration is about 0.39mg/ml (referring to Fig. 2).
Comparative example 3：Not only with lysate goat dairy cream crack but also useless " potassium phosphate buffer solution-is super
Pure water " washs goat dairy butter oil layer
Step 4) is cleaned by ultrasonic the butter oil layer obtained in step 3) with 3 times of volume ultra-pure waters in this comparative example, has cleaned
At upper layer butter oil is taken under the conditions of latter 4 DEG C, which cleans 3 times repeatedly, and the crude extract of butter oil is obtained after centrifugation.Remaining
Step is identical as comparative example 1.It is measured through BCA protein detection kits (sigma companies), the goat dairy milk fat ball film egg of extraction
White concentration is minimum, is 0.059mg/m (referring to Fig. 2).
Comparative example 4：Goat dairy cream is cracked with traditional PMSF lysates
7) this comparative example step replaces the two sulphur Soviet Union in the present invention using traditional phenylmethylsulfonyl fluoride (PMSF) lysate
Sugar alcohol lysate, remaining step are same as Example 1.It is measured through BCA protein detection kits (sigma companies), the mountain of extraction
Sheep breast milk fat globule membrane protein concentration is about 0.38mg/ml (referring to Fig. 2).
Known to being analyzed to embodiment 1 and 1~4 milk fat globule membrane protein separating resulting of comparative example：
1) lactoprotein yield
With the goat dairy butterfat that BCA protein detection kits (sigma companies) measure embodiment 1, comparative example 1~4 is extracted
Fat ball membrane protein concentration (total protein).A concentration of 0.52mg/ml of the milk fat globule membrane protein wherein obtained in embodiment 1,
Higher than the milk fat globule membrane protein concentration that comparative example 1~4 obtains, and 3 gained milk fat globule membrane protein concentration of comparative example is minimum, only
For 0.059mg/ml, illustrate to use " potassium phosphate buffer solution-ultra-pure water " detergent cream fat deposit and two sulphur threoses in embodiment
The addition of alcohol lysate can effectively improve the recovery rate of milk fat globule membrane protein.
2) by a nanoliter tablets by HPLC-MS (nano HPLC-MS/MS) to embodiment 1, comparative example 1-4
Obtained goat dairy milk fat globule membrane protein is analyzed.The milk fat globule membrane protein property that embodiment 1 is extracted is more stable, impurity
Peak area is much smaller than the impurity peak area in comparative example, and Mass Spectrometer Method result shows that the protein quantity in embodiment 1 is about pair
Twice of ratio 3 illustrates that the goat dairy milk fat globule membrane protein purity that embodiment 1 obtains is significantly improved.
Claims (4)
1. a kind of extracting method of goat dairy milk fat globule membrane protein for proteomics research, there is following process steps；
1) Mining takes goat dairy as whole milk samples；
2) sucrose being added in whole milk samples is stirred, and obtains uniform goat dairy sucrose solution；Sucrose addition presses matter
Amount is calculated as the 4% of whole milk samples；
3) goat dairy sucrose solution under the conditions of 4 DEG C is centrifuged into 1h, centrifugal force 50000g, collects upper layer butter oil layer；
4) it uses potassium phosphate buffer solution to be cleaned by ultrasonic butter oil layer, takes upper layer butter oil, it is clear to repeat potassium phosphate buffer solution ultrasound
It washes 1 time；It is cleaned by ultrasonic 1 time with ultra-pure water again, is centrifugally separating to obtain the crude extract of butter oil；Each cleaning potassium phosphate used
Buffer solution, ultra-pure water are 3 times of upper layer butter oil by volume；
5) crude extract of butter oil is stood to butter oil under the conditions of 4 DEG C and is solidified；
6) the butterfat solids heating water bath of solidification is melted into cream；
7) lysate is added into liquid cream and is stored at room temperature 50~60min, lysate addition is the 0.1 of liquid milk oil volume
Times, then the grease on centrifugation removal upper layer, collects lower sediment, obtains the milk fat globule membrane protein solution of goat dairy；Described
Lysate, composition and its mol ratio are dithiothreitol (DTT)s:Tris-HCl:Lauryl sodium sulfate=15:1:60~63, and two
A concentration of 1.5mM of the sulphur threitol in lysate；
8) pre- cold acetone precipitation：Pre- cold acetone mixing is added into milk fat globule membrane protein solution, the temperature of pre- cold acetone is -20
DEG C, dosage is 5 times of milk fat globule membrane protein solution；It is placed 10~15 hours at -20 DEG C；Supernatant is abandoned in centrifugation, then use etc.
The pre- cold acetone of volume is washed 2 times, then sucks supernatant after centrifuging, and is air-dried precipitation and is obtained goat dairy milk fat globule membrane protein.
2. the extracting method of the goat dairy milk fat globule membrane protein according to claim 1 for proteomics research,
It is characterized in that, in step 1), the whole milk samples are the fresh goat dairies that Mining takes；Or -20 in 1~3 hour
The goat dairy transported back at~4 DEG C；Or it is positioned over the goat dairy that placement is stored under -80 DEG C of environment, extract goat dairy milk fat ball film
4 DEG C of refrigerator defrostings are placed it in before albumen.
3. the extracting method of the goat dairy milk fat globule membrane protein according to claim 1 for proteomics research,
It is characterized in that, in step 6), the heating water bath, bath temperature is 40~50 DEG C.
4. the extraction of the goat dairy milk fat globule membrane protein according to claim 1,2 or 3 for proteomics research
Method, which is characterized in that in step 8), the pre- cold acetone mixing of the addition carries out under ice bath；Described air-dries
Precipitation, air-dry time are 5~10min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810163762.1A CN108299552A (en) | 2018-02-27 | 2018-02-27 | The extracting method of goat dairy milk fat globule membrane protein for proteomics research |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810163762.1A CN108299552A (en) | 2018-02-27 | 2018-02-27 | The extracting method of goat dairy milk fat globule membrane protein for proteomics research |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108299552A true CN108299552A (en) | 2018-07-20 |
Family
ID=62848905
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810163762.1A Pending CN108299552A (en) | 2018-02-27 | 2018-02-27 | The extracting method of goat dairy milk fat globule membrane protein for proteomics research |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108299552A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111227039A (en) * | 2018-11-29 | 2020-06-05 | 内蒙古伊利实业集团股份有限公司 | Extraction method of fat globule membrane, yoghourt rich in fat globule membrane and preparation method |
| CN111537744A (en) * | 2020-05-26 | 2020-08-14 | 上海交通大学医学院附属上海儿童医学中心 | ELISA quantitative detection method of milk agglutinin or mucin 1 based on milk fat globule membrane characteristics |
| CN112062809A (en) * | 2020-08-20 | 2020-12-11 | 天津科技大学 | Method for extracting milk fat globules and product prepared by method |
| CN113455551A (en) * | 2021-07-08 | 2021-10-01 | 中国农业大学 | Phospholipid-rich milk fat globule membrane, preparation method thereof and application thereof in promoting brain development and relieving cognitive function decline |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1699414A (en) * | 2005-02-28 | 2005-11-23 | 上海第二医科大学附属新华医院 | Functional peptide of milk agglutinin separated from MCF-7 and method for preparing same |
| JP2007131550A (en) * | 2005-11-08 | 2007-05-31 | Snow Brand Milk Prod Co Ltd | Immunity function regulator |
| CN102863526A (en) * | 2012-09-29 | 2013-01-09 | 哈尔滨工业大学 | Separation method of yak milk milk fat globule membrane protein |
| US20130129889A1 (en) * | 2008-10-08 | 2013-05-23 | Wisconsin Alumni Research Foundation | Method to separate lipids from cheese whey and fat-free whey protein product formed thereby |
-
2018
- 2018-02-27 CN CN201810163762.1A patent/CN108299552A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1699414A (en) * | 2005-02-28 | 2005-11-23 | 上海第二医科大学附属新华医院 | Functional peptide of milk agglutinin separated from MCF-7 and method for preparing same |
| JP2007131550A (en) * | 2005-11-08 | 2007-05-31 | Snow Brand Milk Prod Co Ltd | Immunity function regulator |
| US20130129889A1 (en) * | 2008-10-08 | 2013-05-23 | Wisconsin Alumni Research Foundation | Method to separate lipids from cheese whey and fat-free whey protein product formed thereby |
| CN102863526A (en) * | 2012-09-29 | 2013-01-09 | 哈尔滨工业大学 | Separation method of yak milk milk fat globule membrane protein |
Non-Patent Citations (7)
| Title |
|---|
| TIMOTHY A REINHARDT 等: "Bovine Milk Fat Globule Membrane Proteome", 《JOURNAL OF DAIRY RESEARCH》 * |
| TIMOTHY A. REINHARDT 等: "Ca2+-ATPase protein expression in mammary tissue", 《AM J PHYSIOL CELL PHYSIOL》 * |
| 叶清 等: "人初乳与牛初乳中乳脂肪球膜蛋白质组成的对比研究", 《乳业科学与技术》 * |
| 景萌娜 等: "母乳和牛乳中乳脂肪球膜蛋白质的差异分析", 《食品科学》 * |
| 杨梅 等: "人乳与牛乳乳脂肪球膜蛋白质组的对比研究", 《现代食品科技》 * |
| 赵小伟 等: "牛奶和山羊奶中乳脂球膜蛋白的比较研究", 《中国畜牧兽医》 * |
| 都军霞 等: "乳脂肪球膜主要膜蛋白功能的研究进展", 《中国畜牧兽医》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111227039A (en) * | 2018-11-29 | 2020-06-05 | 内蒙古伊利实业集团股份有限公司 | Extraction method of fat globule membrane, yoghourt rich in fat globule membrane and preparation method |
| CN111537744A (en) * | 2020-05-26 | 2020-08-14 | 上海交通大学医学院附属上海儿童医学中心 | ELISA quantitative detection method of milk agglutinin or mucin 1 based on milk fat globule membrane characteristics |
| CN111537744B (en) * | 2020-05-26 | 2023-04-07 | 上海交通大学医学院附属上海儿童医学中心 | ELISA quantitative detection method of milk agglutinin or mucin 1 based on milk fat globule membrane characteristics |
| CN112062809A (en) * | 2020-08-20 | 2020-12-11 | 天津科技大学 | Method for extracting milk fat globules and product prepared by method |
| CN113455551A (en) * | 2021-07-08 | 2021-10-01 | 中国农业大学 | Phospholipid-rich milk fat globule membrane, preparation method thereof and application thereof in promoting brain development and relieving cognitive function decline |
| CN113455551B (en) * | 2021-07-08 | 2024-03-29 | 中国农业大学 | Phospholipid-rich milk fat globule membrane, preparation method thereof and application thereof in promoting brain development and relieving cognitive decline |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108299552A (en) | The extracting method of goat dairy milk fat globule membrane protein for proteomics research | |
| KR101761680B1 (en) | Isolation Method of Extracellular Vesicles by Aqueous Two-phase System | |
| CN110161147A (en) | The high-throughput quantification measuring method of free oligosaccharides in cream | |
| EP3246703A1 (en) | Method and kit for capturing extracellular vesicles (evs) on a solid surface | |
| CN108680756A (en) | A kind of incomplete antibody detection kit and detection method | |
| CN107840883A (en) | The method that 3 kinds of key components of bovine casein separate simultaneously | |
| Barrett | The serological diagnosis of glandular fever (infectious mononucleosis): a new technique | |
| Schoemig et al. | Solid liquid liquid extraction of porcine gastric mucins from homogenized animal material | |
| CN103602632A (en) | Method for separating fetal nucleated red blood cells in maternal peripheral blood | |
| Zittle et al. | The fat-globule membrane of milk: Alkaline phosphatase and xanthine oxidase in skimmilk and cream | |
| Hughes et al. | Immunological and chemical investigations of vaccine virus: II. chemical analysis of elementary bodies of vaccinia | |
| Svedberg et al. | The molecular weight of casein. II | |
| WO1998026284A1 (en) | Selective lysis of cells | |
| WO2020035033A1 (en) | Biomarker calb1 for diagnosing early stages of diabetic nephropathy | |
| Marks et al. | The enzymatic measurement of glucose by autoanalysis | |
| CN109507127A (en) | It is a kind of for detecting the kit of LBP content in human blood | |
| McPherson et al. | Isolation of bovine milk fat globule membrane material from cream without prior removal of caseins and whey proteins | |
| CN110987717B (en) | Analysis method of yoghourt | |
| US6852540B2 (en) | Control serum for dry analytical element | |
| Wilson et al. | Rapid micro procedure for scanning distribution of subcellular components in density gradient fractions of a tissue homogenate | |
| Monroy et al. | A cytoplasmic fraction acting on the surface layers of the Arbacia punctulata eggs | |
| Galloway et al. | Micro Determination of Cholesterol by Use of 0.04 Ml. of Blood Serum: Adapted from the Method of Sperry and Webb | |
| JP2007139759A (en) | Protein fraction device, and protein fraction and concentration method | |
| Wagstaff et al. | A Simple Routine Radioisotopic Method for the Estimatin of Serum Vitamin B12 Using DEAE Cellulose and Human Serum Binding Agent | |
| JP4276205B2 (en) | Specimen preparation method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20180720 |