CN109856259A - A kind of serum cancer of pancreas excretion body differential protein and its again verification process - Google Patents
A kind of serum cancer of pancreas excretion body differential protein and its again verification process Download PDFInfo
- Publication number
- CN109856259A CN109856259A CN201811643412.1A CN201811643412A CN109856259A CN 109856259 A CN109856259 A CN 109856259A CN 201811643412 A CN201811643412 A CN 201811643412A CN 109856259 A CN109856259 A CN 109856259A
- Authority
- CN
- China
- Prior art keywords
- excretion body
- protein
- albumen
- serum
- pancreas
- 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
Landscapes
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention provides a kind of serum cancer of pancreas excretion body differential protein, including albumin A DAM10, PROTEIN C FHR1, albumin A POA4, albumen Testicular tissue protein Li 70, albumen HEL-S-78p, albumen PGLYRP2, protein I GFALS, protein B CHE, albumin A NPEP, albumen LBP, PROTEIN C 9, albumen PZP, albumin A NXA11.The present invention also provides a kind of serum cancer of pancreas excretion body differential protein verification process again, comprising the following steps: in S1, separation blood serum sample without excretion body serum and excretion body；S2, using the isolated excretion body of step S1, prepare excretion body PBS solution；S3, the ultra microstructure that excretion body is checked by transmission electron microscope；S4, the expression that some excretion body surfaces face marker protein is detected by western blot analysis.Serum cancer of pancreas excretion body differential protein of the invention, can be used in preparing the Related product of diagnosis of pancreatic cancer, serum cancer of pancreas excretion body differential protein again verification process have the advantages that absolute specificity, high throughput, it is noiseless, can identify macromolecular isomers and small molecule.
Description
Technical field
The invention belongs to the verification method technical fields of serum cancer of pancreas excretion body differential protein, and in particular to a kind of serum
Cancer of pancreas excretion body differential protein and its again verification process.
Background technique
Currently, the verification method of serum cancer of pancreas excretion body differential protein is with outside the corresponding detection of specific antibody of albumen
Secrete the method for body.Cell and the method for organizing common Western Blot: it is separated by PAGE (polyacrylamide gel electrophoresis)
Protein example, be transferred on solid phase carrier (such as cellulose nitrate film), with the protein or polypeptide on solid phase carrier
As antigen, immune response is played with corresponding antibody, then react with the secondary antibody of enzyme or isotope labelling, it is aobvious by substrate
Color or autoradiograph are to detect the protein ingredient of the specific destination gene expression of electrophoretic separation.The liquid such as serum are common
The method of ELISA: 1. making antibody be integrated to certain surface of solid phase carriers, and keeps its immunocompetence.2. making antibody and certain enzyme
Enzyme labelled antibody is connected into, this enzyme labelled antibody had not only retained its immunocompetence, but also retained the activity of enzyme.In measurement, being marked by inspection
This and enzyme labelled antibody react by the antibody of different step and surface of solid phase carriers.Make shape on solid phase carrier with the method for washing
At antigen antibody complex and other substances separate, finally combine tested substance in enzyme amount on solid phase carrier and sample
Measure into certain ratio.After the substrate of enzyme reaction is added, substrate becomes color products by enzymatic, in the amount and sample of product by
The amount for examining substance is directly related, therefore can whether there is or not qualitative or quantitative analysis according to the depth of color reaction.However, various albumen need
Corresponding various antibody, and antibody acquisition difficulty is big, quality is also irregular, is one of the technology barrier that research faces.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of serum cancer of pancreas excretion body differential protein, its application and its again
Verification process, to solve the problems, such as proposed in background technique；Simultaneously to pancreatopathy cancer serum excretion body differential protein (ADAM10,
CFHR1、APOA4、Testicular tissue protein Li 70、HEL-S-78p、PGLYRP2、IGFALS、BCHE、
ANPEP, LBP, C9, PZP, ANXA11) it is verified.
In order to solve the above technical problems, the embodiment of the present invention provides a kind of serum cancer of pancreas excretion body differential protein,
It is characterized in that, including albumin A DAM10, PROTEIN C FHR1, albumin A POA4, albumen Testiculartissue protein Li
70, albumen HEL-S-78p, albumen PGLYRP2, protein I GFALS, protein B CHE, albumin A NPEP, albumen LBP, PROTEIN C 9, egg
White PZP, albumin A NXA11.
The present invention also provides a kind of verification process again of serum cancer of pancreas excretion body differential protein, which is characterized in that including
Following steps: S1, separation blood serum sample in without excretion body serum and excretion body；S2, using the isolated excretion of step S1
Body prepares excretion body PBS solution；S3, the ultra microstructure that excretion body is checked by transmission electron microscope；S4, pass through egg
White matter engram analysis detects the expression of some excretion body surfaces face marker protein.
Further, the step S1, separation blood serum sample in without excretion body serum and excretion body, specifically include following
Process: blood serum sample is thawed and is centrifuged 30 minutes at 4 DEG C with small supercentrifuge 10000g, to remove non-in sample
Excretion body microvesicle；To go unless supernatant after excretion body microvesicle with small supercentrifuge 110000g centrifugation 70 minutes after, by nothing
Excretion body serum separate it is to be checked to another Restored in test tube, excretion body precipitating be resuspended in 4ml PBS.
Further, the step S2, prepare excretion body PBS solution, specifically include following procedure: will be obtained by step S1
Excretion weight suspension at 4 DEG C with supercentrifuge 110000g ultracentrifugation 70 minutes small, keep excretion body in suspension heavy
It forms sediment；Final excretion body precipitating is resuspended in the PBS of 200 μ l, excretion body PBS solution is obtained.
Further, the step S4, western blot analysis detect the table of some excretion body surfaces face marker protein
Up to process, comprising the following steps: (1) protein quantification；(2) pancreatin digests；(3) liquid chromatography-mass spectrometry；(4) data
Processing；
Wherein, the process of step (1) protein quantification carries out determination of protein concentration using BCA kit.
Wherein, step (2) the pancreatin enzymatic hydrolysis, specifically includes following procedure: dithiothreitol (DTT) is added in protein solution to be made
Its final concentration of 5mM, 56 DEG C of reduction 30min；Iodo-acetamide is added later makes its final concentration of 11mM, and room temperature is protected from light incubation
15min；Finally the urea concentration of sample is diluted to lower than 2M；With pancreatin: pancreas is added in the mass ratio that the ratio between albumen is 1:50
Enzyme, 37 DEG C of enzymatic hydrolysis are overnight；Again with pancreatin: pancreatin is added in the mass ratio of the ratio between albumen 1:100, continues to digest 4h；
Wherein, step (3) liquid chromatography-mass spectrometry, specifically includes following procedure: a, peptide fragment liquid phase color
It is separated after spectrum mobile phase A phased soln using 1000 ultra high efficiency liquid phase systems of EASY-nLC；Mobile phase A is containing 0.1% formic acid
With the aqueous solution of 2% acetonitrile；Mobile phase B is the aqueous solution containing 0.1% formic acid and 90% acetonitrile；The setting of liquid phase gradient: 0~
40min, 7%-25%B；40~52min, 25%-35%B；52~56min, 35%-80%B；56~60min, 80%B, stream
Speed maintains 450nL/min；B, then peptide fragment is ionized in injection NSI ion source via after the separation of ultra high efficiency liquid phase systems
It is analyzed into Q ExactiveTM Plus mass spectrum；Ion source voltage is set as 2.0kV, peptide fragment parent ion and its secondary fragment
All it is detected and analyzed using high-resolution Orbitrap；First mass spectrometric scanning range is set as 350~950m/z, scanning point
Resolution is set as 70,000；Second order ms Orbitrap scanning resolution is set as 17500；C, data acquisition scheme uses data
The fragmentation energies of independent form scanner program, HCD collision cell are set as 27；First mass spectrometric automatic growth control is set as 3E6, most
The big ion implanting time is set as 50ms；Second order ms automatic growth control is set as 1E5, and maximum ion injection length is set as
150ms, isolation window are set as 1.6m/z；
Wherein, step (4) data processing, specifically include following procedure: peptide fragment parameter: protease is set as
Trypsin, maximum leakage enzyme site number are set as 0, and peptide segment length is set as 7~25 amino acid residues, and cysteine alkane is arranged
Base turns to fixed modification；Transition parameter: parent ion charge is set as 2,3, and daughter ion charge is set as 1, ionic type
It is set as b, y；To last one since third, the matched quality error tolerance of ion is set as fragment ion selection
0.02Da。
The advantageous effects of the above technical solutions of the present invention are as follows: serum cancer of pancreas excretion body differential protein of the invention,
Including albumin A DAM10, PROTEIN C FHR1, albumin A POA4, albumen Testicular tissueprotein Li 70, albumen
HEL-S-78p, albumen PGLYRP2, protein I GFALS, protein B CHE, albumin A NPEP, albumen LBP, PROTEIN C 9, albumen PZP, egg
White ANXA11；By using the verification process again of PRM to test relative to original Western Blot/ELISA in verification process again
Card method, this verification method have absolute specificity, high throughput, it is noiseless, can identify macromolecular isomers and small molecule
The advantages of；And the verification result of verification process is shown again: ADAM10, ANPEP, ANXA11 are higher than in pancreatopathy cancer serum excretion body
Pancreatitis and normal serum excretion body, and three is peculiar in excretion body；Testicular tissue protein Li 70,
HEL-S-78p is significantly higher than cancer of pancreas and pancreatitis serum excretion body in normal serum excretion body；Meanwhile utilization is authenticated again
The effects of journey can play diagnosis to cancer of pancreas and judge by stages.
Detailed description of the invention
Fig. 1 is the differential protein abundance statistical chart in the serum excretion body in the present invention；
Fig. 2 is in the present invention without the differential protein abundance statistical chart in excretion body serum.
Specific embodiment
To keep the technical problem to be solved in the present invention, technical solution and advantage clearer, below in conjunction with attached drawing and tool
Body embodiment is described in detail.
In the description of the present invention, it should be noted that term " center ", "upper" "lower", "left", "right", "vertical",
The orientation or positional relationship of the instructions such as "horizontal", "inner", "outside", "front", "rear" is that orientation based on the figure or position are closed
System, is merely for convenience of description of the present invention and simplification of the description, rather than the device or element of indication or suggestion meaning must have
Specific orientation is constructed and operated in a specific orientation, therefore is not considered as limiting the invention.In addition, term " the
One ", " second ", " third " are used for descriptive purposes only and cannot be understood as indicating or suggesting relative importance.
In the description of the present invention, it should be noted that unless otherwise clearly defined and limited, term " installation ", " phase
Even ", " connection " should be used as broadly understood, for example, it may be being fixedly connected, may be a detachable connection, or be integrally connected；
It can be mechanical connection, be also possible to be electrically connected；It can be directly connected, can also indirectly connected through an intermediary, it can be with
It is the connection inside two elements.For the ordinary skill in the art, it can understand that above-mentioned term exists with concrete condition
Concrete meaning in the present invention.
A kind of serum cancer of pancreas excretion body differential protein, including albumin A DAM10, PROTEIN C FHR1, albumin A POA4, albumen
Testicular tissue protein Li 70, albumen HEL-S-78p, albumen PGLYRP2, protein I GFALS, albumen
BCHE, albumin A NPEP, albumen LBP, PROTEIN C 9, albumen PZP, albumin A NXA11.
Using the initial data of MaxQuant software processing LC-MS/MS, then carried out for mankind Uniprot database
Search.Identify 453 respectively from the serum excretion body that Pancreas cancer patients, Pancreatitis Patients and healthy individuals obtain respectively,
460 and 384 kind of protein.
Protein is compared as follows in the serum excretion body that Pancreas cancer patients, Pancreatitis Patients and healthy individuals obtain:
It come the expression of normalized protein and is analyzed from Pancreas cancer patients, Pancreatitis Patients and health with LFQ algorithm
Significant difference between three groups of serum excretion bodies of body.To at least there are two the data of non-null value to carry out in three repeated experiments data
Statistical analysis, and when meeting following screening criteria, it is believed that the significant difference of the data obtained: multiple > 2.0 of differential expression and
Value < 0.05 P.It is compared with the serum excretion body from healthy individuals, the serum excretion body from Pancreas cancer patients has 29 species diversity
The protein of expression: the up-regulation of 18 kinds of protein (be respectively: FGA, PTTG1IP, ANPEP, LBP, LTF, FGG, HEL-S-78P,
C9, VWF, SLC4A1, FTL, ANXA11, SDCBP, C4B, CD9, F9, IGHA2, VWF), 11 kinds of protein lower (be respectively:
TTR、TUBB1、PZP、PGLYRP2、DKFZp686M0562、IGFALS、BCHE、IGKV3-20、FCN2、DKFZp686O1553、
B4DU16).It is compared with the serum excretion body from pancreatitis, the serum excretion body from Pancreas cancer patients has 28 species diversity tables
The protein reached: the up-regulation of 14 kinds of protein (be respectively: HEL46, FGG, HEL-S-78P, FGA, CFP, APAO4, CD9,
A2JA19, ANXA11, ITGB3, CAMP, TF, CFHR1, ADAM10), 14 kinds of protein lower (be respectively: A2NH55,
DKFZp779K0533、F9、AHSG、SPP2、DEFA3、CRP、PZP、F7、SERPINA10、S100A9、F10、PROC、IGKV3-
20)；Differential protein abundance statistics in serum excretion body is as shown in Figure 1；Differential protein abundance statistics in no excretion body serum
As shown in Figure 2.
A kind of serum cancer of pancreas excretion body differential protein of the invention, can be used for preparing the product of diagnosis of pancreatic cancer.Wherein,
The product is chip, detection reagent or detection kit.
Serum cancer of pancreas excretion body differential protein of the invention, including albumin A DAM10, PROTEIN C FHR1, albumin A POA4,
Albumen Testicular tissue protein Li 70, albumen HEL-S-78p, albumen PGLYRP2, protein I GFALS, albumen
BCHE, albumin A NPEP, albumen LBP, PROTEIN C 9, albumen PZP, albumin A NXA11；And the verification result of verification process is shown again:
ADAM10, ANPEP, ANXA11 are higher than pancreatitis and normal serum excretion body in pancreatopathy cancer serum excretion body, and three is outside
It secretes peculiar in body；Testicular tissue protein Li 70, HEL-S-78p are significantly high in normal serum excretion body
In cancer of pancreas and pancreatitis serum excretion body；Meanwhile diagnosis can be played to cancer of pancreas with verification process again and is judged by stages
The effects of.
A kind of verification process again of serum cancer of pancreas excretion body differential protein, comprising the following steps: S1, separation blood serum sample
In without excretion body serum and excretion body；Blood serum sample is thawed and is centrifuged 30 points at 4 DEG C with small supercentrifuge 10000g
Clock, to remove the non-excretion body microvesicle in sample；It will go unless the supernatant after excretion body microvesicle is with small supercentrifuge
110000g be centrifuged 70 minutes after, will be separated without excretion body serum it is to be checked to another Restored in test tube, excretion body precipitating be resuspended in 4ml
In PBS.S2, using the isolated excretion body of step S1, prepare excretion body PBS solution；By the resulting excretion body of step S1
Re-suspension liquid with supercentrifuge 110000g ultracentrifugation 70 minutes small, precipitates excretion body in suspension at 4 DEG C；It will be final
Excretion body precipitating be resuspended in the PBS of 200 μ l, obtain excretion body PBS solution.S3, it is checked by transmission electron microscope
The ultra microstructure of excretion body；It can be seen that diameter is that the transparent, round or ellipse of 30-100nm has film envelope in TEM experiment
The particle that closed.S4, detected by western blot analysis some excretion body surfaces face marker protein (CD9, CD63 and
TSG101 expression).These labels are significantly expressed in excretion body.
The step S4, western blot analysis detect the expression process of some excretion body surfaces face marker protein, packet
Include following steps:
(1) protein quantification；Determination of protein concentration is carried out using BCA kit.
(2) dithiothreitol (DTT) is added in protein solution makes its final concentration of 5mM, 56 DEG C of reduction 30min；Iodo is added later
Acetamide makes its final concentration of 11mM, and room temperature, which is protected from light, is incubated for 15min；Finally the urea concentration of sample is diluted to lower than 2M；With
Pancreatin: pancreatin is added in the mass ratio that the ratio between albumen is 1:50, and 37 DEG C of enzymatic hydrolysis are overnight；Again with pancreatin: the matter of the ratio between albumen 1:100
Pancreatin is added in amount ratio, continues to digest 4h.
(3) liquid chromatography-mass spectrometry；Liquid chromatography-mass spectrometry specifically includes following procedure: a, peptide
It is separated after section liquid chromatogram mobile phase A phased soln using 1000 ultra high efficiency liquid phase systems of EASY-nLC；Mobile phase A is
Aqueous solution containing 0.1% formic acid and 2% acetonitrile；Mobile phase B is the aqueous solution containing 0.1% formic acid and 90% acetonitrile；Liquid phase gradient
Setting: 0~40min, 7%-25%B；40~52min, 25%-35%B；52~56min, 35%-80%B；56~60min,
80%B, flow velocity maintain 450nL/min；B, peptide fragment is carried out in injection NSI ion source via after the separation of ultra high efficiency liquid phase systems
Then ionization is analyzed into Q ExactiveTM Plus mass spectrum；Ion source voltage is set as 2.0kV, peptide fragment parent ion and its
Secondary fragment is all detected and analyzed using high-resolution Orbitrap；First mass spectrometric scanning range is set as 350~950m/
Z, scanning resolution are set as 70,000；Second order ms Orbitrap scanning resolution is set as 17500；C, data acquisition scheme
Using data independent form scanner program, the fragmentation energies of HCD collision cell are set as 27；The setting of first mass spectrometric automatic growth control
For 3E6, maximum ion injection length is set as 50ms；Second order ms automatic growth control is set as 1E5, when maximum ion is injected
Between be set as 150ms, isolation window is set as 1.6m/z.
(4) data processing；Specifically include following procedure: peptide fragment parameter: protease is set as Trypsin, and position is cut in maximum leakage
Points are set as 0, and peptide segment length is set as 7~25 amino acid residues, and setting cysteine is alkylated to fixed modification；
Transition parameter: parent ion charge is set as 2,3, and daughter ion charge is set as 1, and ionic type is set as b, y；Fragment from
To last one since third, the matched quality error tolerance of ion is set as 0.02Da for son selection.
In the present embodiment, the optional supplier of used reagent is as follows:
In the present invention, the difficulty that the very good solution of verification process again of PRM mass-spectrometric technique antibody obtains is big, quality ginseng
The uneven problem of difference.Below table compares what original (Western Blot/ELISA) verification method was used with the present invention
PRM mass-spectrometric technique verification process again.
The present invention has absolute specificity, height using the verification process again that PRM carries out serum cancer of pancreas excretion body differential protein
Flux, it is noiseless, can identify macromolecular isomers and small molecule, solve antibody acquisition difficulty is big, quality is irregular
The problem of, macromolecular isomers and small molecule can be accurately identified.
The above is a preferred embodiment of the present invention, it is noted that for those skilled in the art
For, without departing from the principles of the present invention, it can also make several improvements and retouch, these improvements and modifications
It should be regarded as protection scope of the present invention.
Claims (5)
1. a kind of serum cancer of pancreas excretion body differential protein, which is characterized in that including albumin A DAM10, PROTEIN C FHR1, albumen
APOA4, albumen Testicular tissue protein Li 70, albumen HEL-S-78p, albumen PGLYRP2, albumen
IGFALS, protein B CHE, albumin A NPEP, albumen LBP, PROTEIN C 9, albumen PZP, albumin A NXA11.
2. a kind of verification process again of serum cancer of pancreas excretion body differential protein according to claim 1, which is characterized in that
The following steps are included: S1, separation blood serum sample in without excretion body serum and excretion body；It is S2, isolated using step S1
Excretion body prepares excretion body PBS solution；S3, the ultra microstructure that excretion body is checked by transmission electron microscope；S4, lead to
Western blot analysis is crossed to detect the expression of some excretion body surfaces face marker protein.
3. a kind of verification process again of serum cancer of pancreas excretion body differential protein according to claim 2, which is characterized in that
The step S1, separation blood serum sample in without excretion body serum and excretion body, specifically include following procedure: by blood serum sample solution
Freeze and at 4 DEG C with small supercentrifuge 10000g centrifugation 30 minutes, to remove the non-excretion body microvesicle in sample；It will removal
After supernatant after non-excretion body microvesicle was with small supercentrifuge 110000g centrifugation 70 minutes, will be separated without excretion body serum to
Another Restored in test tube is to be checked, and excretion body precipitating is resuspended in 4ml PBS.
4. a kind of verification process again of serum cancer of pancreas excretion body differential protein according to claim 2, which is characterized in that
The step S2, excretion body PBS solution is prepared, specifically includes following procedure: by the resulting excretion weight suspension of step S1 4
With supercentrifuge 110000g ultracentrifugation 70 minutes small at DEG C, precipitate excretion body in suspension；Final excretion body is sunk
Shallow lake is resuspended in the PBS of 200 μ l, obtains excretion body PBS solution.
5. a kind of verification process again of serum cancer of pancreas excretion body differential protein according to claim 2, which is characterized in that
The step S4, western blot analysis detect the expression process of some excretion body surfaces face marker protein, including following step
It is rapid: (1) protein quantification；(2) pancreatin digests；(3) liquid chromatography-mass spectrometry；(4) data processing；
Wherein, the process of step (1) protein quantification carries out determination of protein concentration using BCA kit；
Wherein, step (2) the pancreatin enzymatic hydrolysis, specifically includes following procedure: dithiothreitol (DTT) is added in protein solution makes its end
Concentration is 5mM, 56 DEG C of reduction 30min；Iodo-acetamide is added later makes its final concentration of 11mM, and room temperature, which is protected from light, is incubated for 15min；
Finally the urea concentration of sample is diluted to lower than 2M；With pancreatin: the mass ratio addition pancreatin that the ratio between albumen is 1:50,37 DEG C
Enzymatic hydrolysis is overnight；Again with pancreatin: pancreatin is added in the mass ratio of the ratio between albumen 1:100, continues to digest 4h；
Wherein, step (3) liquid chromatography-mass spectrometry, specifically includes following procedure: a, peptide fragment liquid chromatogram stream
It is separated after dynamic phase A phased soln using 1000 ultra high efficiency liquid phase systems of EASY-nLC；Mobile phase A be containing 0.1% formic acid and
The aqueous solution of 2% acetonitrile；Mobile phase B is the aqueous solution containing 0.1% formic acid and 90% acetonitrile；The setting of liquid phase gradient: 0~40min,
7%-25%B；40~52min, 25%-35%B；52~56min, 35%-80%B；56~60min, 80%B, flow velocity maintain
In 450nL/min；B, then peptide fragment ionize in injection NSI ion source into Q via after the separation of ultra high efficiency liquid phase systems
ExactiveTM Plus mass spectrum is analyzed；Ion source voltage is set as 2.0kV, and peptide fragment parent ion and its secondary fragment all make
It is detected and analyzed with high-resolution Orbitrap；First mass spectrometric scanning range is set as 350~950m/z, scanning resolution
It is set as 70,000；Second order ms Orbitrap scanning resolution is set as 17500；C, data acquisition scheme using data it is non-according to
Rely type scanner program, the fragmentation energies of HCD collision cell are set as 27；First mass spectrometric automatic growth control is set as 3E6, it is maximum from
Sub- injection length is set as 50ms；Second order ms automatic growth control is set as 1E5, and maximum ion injection length is set as
150ms, isolation window are set as 1.6m/z；
Wherein, step (4) data processing, specifically include following procedure: peptide fragment parameter: protease is set as Trypsin, most
Big leakage enzyme site number is set as 0, and peptide segment length is set as 7~25 amino acid residues, and setting cysteine is alkylated to fix
Modification；Transition parameter: parent ion charge is set as 2,3, and daughter ion charge is set as 1, and ionic type is set as b, y；
To last one since third, the matched quality error tolerance of ion is set as 0.02Da for fragment ion selection.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811643412.1A CN109856259A (en) | 2018-12-29 | 2018-12-29 | A kind of serum cancer of pancreas excretion body differential protein and its again verification process |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811643412.1A CN109856259A (en) | 2018-12-29 | 2018-12-29 | A kind of serum cancer of pancreas excretion body differential protein and its again verification process |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109856259A true CN109856259A (en) | 2019-06-07 |
Family
ID=66893374
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811643412.1A Pending CN109856259A (en) | 2018-12-29 | 2018-12-29 | A kind of serum cancer of pancreas excretion body differential protein and its again verification process |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109856259A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110596403A (en) * | 2019-09-17 | 2019-12-20 | 中山大学附属第一医院 | Specific protein marker for ovarian cancer diagnosis and preparation method thereof |
| CN111323589A (en) * | 2020-03-03 | 2020-06-23 | 南通大学附属医院 | Application of serum exosome ANXA11 in diagnosis of pancreatic cancer |
| CN111366654A (en) * | 2020-04-01 | 2020-07-03 | 上海中科新生命生物科技有限公司 | Protein component analysis method for enriching blood exosomes based on differential centrifugation |
| CN112433054A (en) * | 2020-11-20 | 2021-03-02 | 四川大学华西医院 | Application of C9 protein detection reagent in preparation of lung cancer screening kit |
| CN112481368A (en) * | 2020-11-25 | 2021-03-12 | 中国人民解放军空军军医大学 | Drug eruption plasma exosome protein and kit thereof |
| WO2021174995A1 (en) * | 2020-03-02 | 2021-09-10 | 南通大学附属医院 | Use of jq-1 in preparation of medicine for treating pancreatic cancer and method for verifying inhibition of jq-1 on exosome secretion of pancreatic cancer |
| CN113430267A (en) * | 2021-06-29 | 2021-09-24 | 复旦大学附属中山医院 | Application of chemotherapy-related gene expression characteristics in prediction of pancreatic cancer prognosis |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EA003235B1 (en) * | 1999-01-27 | 2003-02-27 | Эносис, Инк. | Method for preparing membrane vesicles |
| US20110195426A1 (en) * | 2009-07-16 | 2011-08-11 | The General Hospital Corporation | Nucleic acids analysis |
| CN105467118A (en) * | 2015-08-30 | 2016-04-06 | 李小彦 | Pancreatic cancer tumor marker and application method thereof |
| CN105891314A (en) * | 2016-03-09 | 2016-08-24 | 上海中科新生命生物科技有限公司 | Method for screening myocardial infarction disease expression protein through iTRAQ technology |
| CN106770872A (en) * | 2017-01-13 | 2017-05-31 | 中国农业科学院北京畜牧兽医研究所 | A kind of authentication method for serum of broilers protein group |
| CN106950374A (en) * | 2017-04-10 | 2017-07-14 | 南通大学附属医院 | Application of the albumen of Glypican 1 in diagnosis of pancreatic cancer, the detection method of positive excretion bulk concentration and application thereof |
| CN108168977A (en) * | 2018-01-08 | 2018-06-15 | 南通大学附属医院 | Extraction, detection method and the kit of excretion body biomarker MMP-13 |
-
2018
- 2018-12-29 CN CN201811643412.1A patent/CN109856259A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EA003235B1 (en) * | 1999-01-27 | 2003-02-27 | Эносис, Инк. | Method for preparing membrane vesicles |
| US20110195426A1 (en) * | 2009-07-16 | 2011-08-11 | The General Hospital Corporation | Nucleic acids analysis |
| CN105467118A (en) * | 2015-08-30 | 2016-04-06 | 李小彦 | Pancreatic cancer tumor marker and application method thereof |
| CN105891314A (en) * | 2016-03-09 | 2016-08-24 | 上海中科新生命生物科技有限公司 | Method for screening myocardial infarction disease expression protein through iTRAQ technology |
| CN106770872A (en) * | 2017-01-13 | 2017-05-31 | 中国农业科学院北京畜牧兽医研究所 | A kind of authentication method for serum of broilers protein group |
| CN106950374A (en) * | 2017-04-10 | 2017-07-14 | 南通大学附属医院 | Application of the albumen of Glypican 1 in diagnosis of pancreatic cancer, the detection method of positive excretion bulk concentration and application thereof |
| CN108168977A (en) * | 2018-01-08 | 2018-06-15 | 南通大学附属医院 | Extraction, detection method and the kit of excretion body biomarker MMP-13 |
Non-Patent Citations (2)
| Title |
|---|
| ，YU-J. JIAO ET AL.: "Characterization and proteomic profiling of pancreatic cancer‐derived serum exosomes", 《JOURNAL OF CELLULAR BIOCHEMISTRY》 * |
| 李琳 等: "基于串联质谱标签法和平行反应监测技术的氟喹诺酮耐药沙门菌蛋白质组学分析", 《微生物学通报》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110596403A (en) * | 2019-09-17 | 2019-12-20 | 中山大学附属第一医院 | Specific protein marker for ovarian cancer diagnosis and preparation method thereof |
| CN110596403B (en) * | 2019-09-17 | 2022-12-16 | 中山大学附属第一医院 | Specific protein marker for ovarian cancer diagnosis and preparation method thereof |
| WO2021174995A1 (en) * | 2020-03-02 | 2021-09-10 | 南通大学附属医院 | Use of jq-1 in preparation of medicine for treating pancreatic cancer and method for verifying inhibition of jq-1 on exosome secretion of pancreatic cancer |
| CN111323589A (en) * | 2020-03-03 | 2020-06-23 | 南通大学附属医院 | Application of serum exosome ANXA11 in diagnosis of pancreatic cancer |
| CN111366654A (en) * | 2020-04-01 | 2020-07-03 | 上海中科新生命生物科技有限公司 | Protein component analysis method for enriching blood exosomes based on differential centrifugation |
| CN112433054A (en) * | 2020-11-20 | 2021-03-02 | 四川大学华西医院 | Application of C9 protein detection reagent in preparation of lung cancer screening kit |
| CN112481368A (en) * | 2020-11-25 | 2021-03-12 | 中国人民解放军空军军医大学 | Drug eruption plasma exosome protein and kit thereof |
| CN113430267A (en) * | 2021-06-29 | 2021-09-24 | 复旦大学附属中山医院 | Application of chemotherapy-related gene expression characteristics in prediction of pancreatic cancer prognosis |
| CN113430267B (en) * | 2021-06-29 | 2023-03-10 | 复旦大学附属中山医院 | Application of chemotherapy-related gene expression characteristics in prediction of pancreatic cancer prognosis |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109856259A (en) | A kind of serum cancer of pancreas excretion body differential protein and its again verification process | |
| Tanca et al. | Comparison of detergent‐based sample preparation workflows for LTQ‐O rbitrap analysis of the E scherichia coli proteome | |
| Chua et al. | Honey protein extraction and determination by mass spectrometry | |
| Murad et al. | Detection and expression analysis of recombinant proteins in plant‐derived complex mixtures using nanoUPLC‐MSE | |
| Winkler et al. | Isolation and mass spectrometry of transcription factor complexes | |
| Rebecchi et al. | Label-free quantitation: a new glycoproteomics approach | |
| Tsai et al. | Mass spectrometry-based strategies for protein disulfide bond identification | |
| US20200333353A1 (en) | Identification of host cell proteins | |
| WO2016143227A1 (en) | Kit for preparing sample for detecting monoclonal antibody | |
| US11835434B2 (en) | Methods for absolute quantification of low-abundance polypeptides using mass spectrometry | |
| Danquah et al. | Intact Transition Epitope Mapping–Targeted High-Energy Rupture of Extracted Epitopes (ITEM-THREE)*[S] | |
| Zhao et al. | imFASP: an integrated approach combining in-situ filter-aided sample pretreatment with microwave-assisted protein digestion for fast and efficient proteome sample preparation | |
| CN102492017B (en) | Serum polypeptide and mass-spectra semiquantitative method taking same as quantitative reference substance | |
| Luo et al. | Resolving the composition of protein complexes using a MALDI LTQ Orbitrap | |
| Downard | Indirect study of non‐covalent protein complexes by MALDI mass spectrometry: Origins, advantages, and applications of the “intensity‐fading” approach | |
| Alvarado et al. | A comparative study of in-gel digestions using microwave and pressure-accelerated technologies | |
| Drabik et al. | Application of the ETD/PTR reactions in top‐down proteomics as a faster alternative to bottom‐up nanoLC‐MS/MS protein identification | |
| Yamaguchi et al. | Specific isolation of N‐terminal fragments from proteins and their high‐fidelity de novo sequencing | |
| Mao et al. | Lys‐Sequencer: An algorithm for de novo sequencing of peptides by paired single residue transposed Lys‐C and Lys‐N digestion coupled with high‐resolution mass spectrometry | |
| Cruz Villarreal et al. | MIMAS: microfluidic platform in tandem with MALDI mass spectrometry for protein quantification from small cell ensembles | |
| Marie et al. | Native Capillary Electrophoresis–Mass Spectrometry of Near 1 MDa Non‐Covalent GroEL/GroES/Substrate Protein Complexes | |
| Bergquist et al. | Future aspects of psychoneuroimmunology–lymphocyte peptides reflecting psychiatric disorders studied by mass spectrometry | |
| Tummala et al. | Effect of sodium dodecyl sulfate micelles on peptide mass fingerprinting by matrix‐assisted laser desorption/ionization mass spectrometry | |
| Bunkenborg et al. | Covalent perturbation as a tool for validation of identifications and PTM mapping applied to bovine alpha‐crystallin | |
| JP2017161447A (en) | Analysis 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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190607 |
|
| RJ01 | Rejection of invention patent application after publication |