CN101440121A - System for separating protein polypeptide in blood serum and reagent kit thereof - Google Patents
System for separating protein polypeptide in blood serum and reagent kit thereof Download PDFInfo
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- CN101440121A CN101440121A CNA2008101879684A CN200810187968A CN101440121A CN 101440121 A CN101440121 A CN 101440121A CN A2008101879684 A CNA2008101879684 A CN A2008101879684A CN 200810187968 A CN200810187968 A CN 200810187968A CN 101440121 A CN101440121 A CN 101440121A
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Abstract
The invention provides an SPE-C system suitable for separating protein polypeptide from blood serum, an SPE-C reagent kit thereof and usage of the SPE-C system or the SPE-C reagent kit for separating protein polypeptide from blood serum. The steps are: treating SPE-C magnetic beads in the reagent kit in a balanced way, performing magnetic separation and discarding supernatant; putting the SPE-C magnetic beads in a combination buffer for a specific combination with protein and polypeptide in the blood serum, performing magnetic separation and discarding supernatant; adding a cleaning buffer solution to clean the magnetic beads and remove specific combined impurities; adding elution buffer to elute specific protein and polypeptide in the blood serum, and testing the concentration of the polypeptide; and the elution buffer can be used for mass spectrometry analysis directly or be frozen at 20 DEG C below zero for mass spectrometry analysis within 24 hours. The SPE-C system or SPE-C reagent kit has the capability of capturing more micro molecular and low content proteins, and has the advantages of high flux, low cost, rapidness, simplification, flexible operation, less sample consumption, and wide application to the screening and diagnosis of various malignancy markers.
Description
Technical field
The invention belongs to biological technical field, relate to a kind of SPE-C system of protein polypeptide in the separation of serum and SPE-C test kit of preparation thereof of being suitable for, be used for the purposes of separation of serum protein polypeptide more specifically to SPE-C system or SPE-C test kit.
Background technology
Protein is the biomass cells various metabolism of depending on for existence and the main executive of regulatory pathway, and the generation of disease may be relevant with the change of protein modified state, the research of proteomics directly is positioned protein level, but it is different with the research of protein chemistry in the past, the research object of protein group is not the albumen of single or minority, it is focused on is from comprehensively, whole, dynamically, quantitative angle goes to study the function of gene, panorama type sees thoroughly that cell protein is expressed and changes on the integral level that the research method of utilizing protein group is expressed from cell protein, the biological characteristics of cell will more be helped to understand, so utilize the essence of the technical study disease of proteomics more problem can be described than the research on genomic level.Proteomic techniques is studied aspects such as the propagation, differentiation of cell, unusual conversion, tumour form again and has been carried out strong exploration, for the judgement of discovery, curative effect and the prognosis of the early diagnosis of disease, medicine target provides important basis.Many medicines target molecules that plays a role is a protein, and drug development is candidate's target based on the protein expression that specificity in the disease generating process raises the downward modulation of living generally.Utilize the new medicine target of generation, development mechanism, evaluation of proteomics method study of disease, and new biomarker, and be used to instruct clinical trial.This not only brings new thinking, strategy for the treatment various diseases, and is containing huge market outlook.
The development of proteomics also provides new idea and method for new drug research, clinical diagnosis, has produced the notion of " clinical proteomics " thus.The major objective of " clinical proteomics " be exactly utilize that mass-spectrometric technique in the proteomics is highly sensitive, high resolution, advantage is sought the protein that can be used in one group of polypeptide that disease forecasting, diagnosis and treatment detect etc. fast and accurately." clinical proteomics " is the important component part of present proteomics, is the subject that application prospect is arranged most emerging in the proteomics.The purpose of clinical proteomics is exactly to set up new have highly sensitive, high narrow spectrum reliable disease early detection method.
The same with the broad sense proteomics, the major technique of clinical proteome research also comprises 3 aspects: the technology of preparing of separation of (1) serum sample and mass-producing; (2) high-throughput, highly sensitive and high resolution protein analysis and authenticate technology; (3) information biology and statistical analysis technology.
The sample preparation technology of clinical protein group is the key constraints that present clinical proteome research reaches highly sensitive, the mass-producing of high specificity box.Because serum sample is very complicated; contain various chaff interferences such as organic micromolecule compound and mass-producing and salt etc. and thousands of protein and polypeptide in the sample; the difference of protein and polypeptide amount is highly significant also, how can just can detect that representative albumen and polypeptide just become very crucial problem in the serum by simple processing.
Magnetic bead is developed recently a kind of new multifunctional reagent that get up and that be widely used in biomedical sector.The structure of magnetic bead is made up of the polymer shell two portions with magnetic kernel and nuclear external parcel usually.Because magnetic is checked the response of foreign field, magnetic bead can be directed mobile in magnetic field.Utilize this character to position, or magnetic bead is separated rapidly from surrounding medium magnetic bead.The surperficial diversity of polymer shell determined magnetic bead can with various biologically active substances (as antibody, antigen, acceptor, enzyme, nucleic acid etc.) coupling, then can in reaction medium, further discern corresponding antigen or antibody, part after these biologically active substances are fixed on the magnetic bead.Substrate, nucleic acid, thus reach the purpose of separating or detecting.Therefore, magnetic bead integrates function vector and separation function, utilizes physics, chemistry and biomedical principle, and many loaded down with trivial details complicated operations are oversimplified.The cycle of traditional test is shortened greatly, obtained in many fields such as cytology, immunology, microbiology, molecular biology and clinical diagnosis and treatments using widely.
Be that solid-phase media is purified to protein and successfully the range protein in lysate, blood plasma, protoplasma and the ascites carried out separation and purification with the magnetic microsphere.The SPE-C magnetic bead is based on weak positive ion-exchange absorption principle, realizes that under specific buffer system and pH condition magnetic bead is attached with parsing to the absorption of differential protein polypeptide.SPE-C belongs to the magnetic bead of weak positive example exchange principle, and this process is a reversing process.Proteinic SPE-C magnetic bead ion exchange process is divided into two stages: adsorption and desorption is attached.Adsorption process wherein: proteic iso-electric point is greater than ambient condition (AS) at least during 1pH, the SPE-C magnetic bead could be more effective conjugated protein, and proteic iso-electric point and ambient condition (pH) differ big more, and the albumen positive charge quantity of electric charge is many more, and be big more with SPE-C magnetic bead binding capacity; Desorption process wherein: ambient condition (pH) is during greater than proteic iso-electric point at least during 1pH, the albumen of SPE-C magnetic bead absorption just can lose electric charge, become negative charge, albumen could dissociate with the SPE-C magnetic bead, and proteic iso-electric point and ambient condition (pH) differ big more, the albumen negative charge amount is many more, and is dissociated abundant more with the SPE-C magnetic bead.So the condition of buffer medium (pH) is very big in conjunction with how many influences of the kind of specific protein polypeptide and amount for magnetic bead.
Although it is attached with parsing to the absorption of differential protein polypeptide that described system can realize magnetic bead, but still can not satisfy magnetic bead simultaneously to the wide spectrum absorption of protein polypeptide in the serum and the requirement of special absorption.Therefore, if select to be suitable for the magnetic bead Laemmli buffer system Laemmli attached with parsing, become the present problem that needs solution to the absorption of differential protein polypeptide.
Summary of the invention
In order to overcome the shortcoming of prior art, based on the present mass spectroscopy and an urgent demand of clinical proteomics, the contriver is by selecting the ambient condition (pH) of some row, therefrom select the maximum and conjugated protein kind of present literature survey, the highest ambient condition of conjugated protein content, thereby provide SPE-C to be used for separation of serum protein polypeptide system and test kit thereof.Wherein, the SPE-C test kit utilizes the principle of weak positive ion-exchange absorption, cooperate a cover to be suitable for the buffered soln acquisition polypeptide and the protein information from serum as much as possible of mass spectroscopy, utilize the magnetic of SPE-C magnetic bead to separate, and when analyze by mass spectrum, thereby seek differential protein.
Therefore, first purpose of the present invention provides a kind of SPE-C system that is suitable for protein polypeptide in the separation of serum, comprise the level pad, binding buffer liquid, cleaning buffer solution, the elutriant that are applicable to the SPE-C magnetic bead, wherein elutriant is the TFA of 0.1%-3%, and level pad, binding buffer liquid and cleaning buffer solution group are selected from:
0.001-2M pH is the Tris-HCL buffered soln of 7.0-9.0; Or
0.001-2M pH is the carbonate buffer solution of 9.0-11.0; Or
0.001-2M pH is the acetate buffer solution of 2.0-6.0; Or
0.001-2M pH is the glycine buffer of 2.0-6.0; Or
0.001-2M pH is the O-phthalic acid buffer of 2.0-6.0; Or
0.001-2M pH is the citrate buffer solution of 2.0-7.0; Or
0.001-2M pH is the TBS buffered soln of 7.0-9.0; Or
0.001-2M pH is the TE buffered soln of 2.0-7.0; Or
0.001-2M pH is the STE buffered soln of 7.0-9.0; Or
0.001-2M pH is the PBS buffered soln of 2.0-8.0; Or
0.001-2M pH is the SSC buffered soln of 7.0-9.0; Or
0.001-2M pH is the SSPE buffered soln of 7.0-9.0.
In a specific embodiments, level pad is selected from 0.25M, the Citrate trianion of pH2-6, PBS, acetate, glycine, phthalic acid or TE buffered soln;
Binding buffer liquid is selected from 0.15M, the Citrate trianion of pH2-6, PBS, acetate, glycine, phthalic acid or TE buffered soln;
Cleaning buffer solution is selected from 0.35M, the Citrate trianion of pH2-6, PBS, acetate, glycine, phthalic acid or TE buffered soln.
Second purpose of the present invention provides the prepared SPE-C test kit that is used for the separation of serum protein polypeptide of above-mentioned SPE-C system.
In a specific embodiments, described SPE-C test kit comprises the magnetic bead that is used in conjunction with the separation of serum protein polypeptide, wherein magnetic bead SPE-C magnetic bead preferably.
The 3rd purpose of the present invention provides SPE-C system or test kit, is used for the purposes of separation of serum protein polypeptide.
In a specific embodiments, described purposes comprises that step is as follows:
(1) with the SPE-C magnetic bead Balance Treatment in the test kit, magnetic resolution is abandoned supernatant;
(2) the SPE-C magnetic bead combines with albumen, polypeptide generation specificity in the serum in binding buffer liquid, and magnetic resolution is abandoned supernatant;
(3) add cleaning buffered soln and clean magnetic bead, remove the anisogamy impurity of dtex;
(4) add elute soln, specific albumen and polypeptide in the wash-out serum, and measure peptide concentration;
(5) elutriant can be used for directly carrying out mass spectroscopy or frozenly carry out mass spectroscopy within-20 ℃, 24 hours.
In step 5), this purposes is to be used to seek the differential protein relevant with disease, sets up more stable diagnostic model.
The SPE-C magnetic bead be with ferric oxide or Z 250 as magnetic kernel, pan coating one deck poly material synthetic nucleocapsid structure high molecule magnetic microsphere.Various factorss such as production operation by control criterionization have guaranteed that this polymer microsphere SPE-C magnetic bead has that monodispersity is good, homogeneous, various advantages that magnetic content is high, the height homogeneous of physics and chemical characteristics quality.Repeatability between single batch (generally within 5%) has guaranteed the quality and the circulation ratio of experimental result.
Wherein, when seeking suitable ambient condition, we with the basic parameter of the protein polypeptide mass spectrum of this SPE-C magnetic bead kit extraction as Quality Control.Setting under the unified signal to noise ratio condition, on behalf of magnetic bead kit, the basic parameter of total peak number amount, peak area, peak height etc. grasp the ability and the mass spectral quality of protein and polypeptide in the mass spectrum of the protein polypeptide that the SPE-C magnetic bead kit is extracted.Ambient condition is suitable, and in the protein polypeptide mass spectrum of extraction, the peak number order can be many more, represents magnetic bead can filter out more protein from sample, provides bigger possibility for seeking differential protein; Peak area is more little, represents the tolerance range of protein peak high more, and the accuracy of later stage searching differential protein and proteinic evaluation just may be high more; The analysis that combines with signal to noise ratio of overall peak heights, noise cancellation signal is low, and the peak heights height shows that the mass spectrum quality is good, and the result is credible relatively.The 2nd, the ability of consistence, searching difference in the mass spectrum group.
In a specific embodiments, described Quality Control is: setting under the unified signal to noise ratio condition, on behalf of magnetic bead kit, the basic parameter of total peak number amount, peak area, peak height etc. grasp the ability and the mass spectral quality of protein and polypeptide in the mass spectrum of the protein polypeptide that magnetic bead kit is extracted.On behalf of magnetic bead, the peak number order to filter out more protein from sample more, provides bigger possibility for seeking differential protein; The peak area tolerance range of small protein mass peak more is high more, and the later stage is sought the accuracy of differential protein and proteinic evaluation just may be high more; The analysis that combines with signal to noise ratio of overall peak heights, noise cancellation signal is low, and the peak heights height shows that the mass spectrum quality is good, and the result is credible relatively.The 2nd, the ability of consistence (CV＜30%), searching difference in the mass spectrum group.
Beneficial effect
1. SPE-C of the present invention system, compare with the Laemmli buffer system Laemmli of routine, having has higher capture ability to small molecular protein and low kurtosis albumen, and high-throughput, with low cost, fast and simple, flexible operation, amount of samples are few, are used for the screening of multiple malignant tumour mark and the advantage of diagnosis in a large number.
2. the SPE-C test kit among the present invention has very excellent performance, and it can extract abundant protein and polypeptide from serum, and sepn process is simple, and operation is convenient.
3. utilize SPE-C magnetic bead kit of the present invention, extract the protein polypeptide in the serum, obtain all proteic mass spectrums, the mass spectrum difference that compares patient or normal healthy controls by software, obtain the otherness mass spectrum expression pattern of the two, the attribute that is used further to analyze unknown sample carries out the diagnosis of disease.
4. utilize SPE-C magnetic bead kit of the present invention, can make the mass spectroscopy of sample reach high-throughput, highly sensitive better plays a role biological mass spectrometry in clinical proteomics research.
Description of drawings
Fig. 1: the procedure chart of SPE-C bead chromatograph, wherein protein polypeptide is adsorbed onto on the SPE-C magnetic bead in the serum, uses the procedure chart of magnetic resolution flush away impurity.
Fig. 2: the protein polypeptide mass spectrum in normal people's pooled serum that wherein a collection of SPE-C magnetic bead kit is extracted for three times among the present invention.
Embodiment
Below in conjunction with specific examples the present invention is done further introduction in detail.
Embodiment 1: the preparation of SPE-C system of the present invention and SPE-C test kit thereof
According to the magnetic bead system of protein isolate polypeptide, preparation elutriant, level pad, binding buffer liquid and cleaning buffer solution obtain the A-C of system.
| Combination | Level pad 0.25M, pH6.0 | Binding buffer liquid 0.15M, pH5.6 | Cleaning buffer solution 0.35M, pH5.0 |
| System A | Citrate trianion | Phthalic acid | Citrate trianion |
| System B | Acetate | Acetate | Citrate trianion |
| System C | PBS | Acetate | Citrate trianion |
| Contrast D | TE | Glycine | PBS |
Table 1
According to the combination of table 1, cooperate independent 1 bottle of magnetic bead, we obtain test kit A, B, C, D (contrast) respectively.Wherein among the test kit A, SPE-C magnetic bead and equalizing and buffering solution 0.25M, the Citrate trianion of pH6.0, binding buffer liquid 0.15M, pH5.6 phthalic acid, cleaning buffer solution 0.35M, pH5.0 Citrate trianion.Wherein among the test kit B, SPE-C magnetic bead and equalizing and buffering solution 0.25M, the acetate of pH6.0, binding buffer liquid 0.15M, pH5.6 acetate, cleaning buffer solution 0.35M, pH5.0 Citrate trianion.Wherein among the test kit C, SPE-C magnetic bead and equalizing and buffering solution 0.25M, the PBS of pH6.0, binding buffer liquid 0.15M, pH5.6 acetate, cleaning buffer solution 0.35M, pH5.0 Citrate trianion.Wherein among the test kit D, SPE-C magnetic bead and equalizing and buffering solution 0.25M, the TE of pH6.0, binding buffer liquid 0.15M, pH5.6 glycine, cleaning buffer solution 0.35M, pH5.0PBS.
Embodiment 2: SPE-C test kit of the present invention is in conjunction with the process of polypeptide in the serum.
Take out above-mentioned test kit A-D at 4 ℃ of refrigerators, and SPE-C bead suspension one is wherein managed, manually turned upside down mixes bead suspension.Getting 200 μ l, eight platoon sample hoses places on the orifice plate, add 100 μ lSPE-C magnetic beads, sample hose is placed on the magnetic bead separator, leave standstill so that the SPE-C magnetic bead is adherent, with the liquid separation that suspends, liquid is limpid, removes the liquid that suspends with the suction of application of sample rifle, the rifle head should be avoided touching the SPE-C magnetic bead, avoids siphoning away the SPE-C magnetic bead.
Sample hose is put on the orifice plate, add 50 μ lSPE-C equalizing and buffering solution, suction is beaten and is mixed, and mixing 18min under 30 ℃ of conditions is placed on sample hose on the magnetic bead separator, leave standstill so that magnetic bead is adherent, with the liquid separation that suspends, liquid is limpid, removes the liquid that suspends with the suction of application of sample rifle, the rifle head should avoid touching magnetic bead, avoids siphoning away magnetic bead.Repeat in the above-mentioned steps twice, guarantee that suspension is sucked away fully.
Sample hose is positioned on the orifice plate, add 80 μ lSPE-C binding buffer solution and 4 μ l serum, inhale repeatedly to beat with the application of sample rifle and make magnetic bead and binding buffer solution suspendible even, suction is played the process strictness and is avoided bubbling, and leaves standstill 30min under 35 ℃ of conditions, comb is positioned over leaves standstill on the magnetic bead separator to the liquid separation of magnetic bead and suspension, liquid is limpid, inhale with the application of sample rifle and to remove the liquid that suspends, the rifle head should avoid touching magnetic bead, avoids siphoning away magnetic bead.
Embodiment 3: test kit cleans magnetic bead and polypeptide conjugates
Sample hose is positioned on the orifice plate, add 80 μ lSPE-C and clean buffered soln, suction is beaten and is mixed, and puts and leaves standstill 5 minutes on the orifice plate, comb is placed on the magnetic bead separator leave standstill, magnetic bead is adherent, with the liquid separation that suspends, liquid is limpid, removes the liquid that suspends with the suction of application of sample rifle, the rifle head should avoid touching magnetic bead, avoids siphoning away magnetic bead.Repeat (above-mentioned) step twice, guarantee that suspension is sucked away fully.
The concentration of polypeptide and mensuration polypeptide in the embodiment 4:SPE-C test kit wash-out serum
Sample hose is positioned on the orifice plate, adds 20 μ l 2.5%TFA elute solns, inhale repeatedly and make a call to more than 10 times, make magnetic bead and elutriant suspendible even, the strictness of piping and druming process is avoided bubbling, and room temperature leaves standstill 3min.
Sample hose is positioned on the magnetic bead separator, left standstill 20 seconds, magnetic bead is fully separated with suspension, supernatant liquor (elutriant) is moved into clean 0.5ml sample hose.(see figure 1) is drawn the part elutriant, measures peptide concentration.
Elutriant can be used for directly carrying out mass spectroscopy or frozenly carry out mass spectroscopy in-20 ℃ within 24 hours.(see figure 2)
Mass spectrometry results (table 2):
| Combination | The peak number order | Repeatability (CV＜30%) |
| Test kit A | 63 | 44/63 |
| Test kit B | 61 | 47/61 |
| Test kit C | 63 | 42/63 |
| Contrast agents box D | 42 | 30/42 |
Wherein, when seeking suitable ambient condition, the basic parameter of the protein polypeptide mass spectrum that our usefulness SPE-C magnetic bead kit is extracted is as Quality Control.Setting under the unified signal to noise ratio condition, on behalf of magnetic bead kit, the basic parameter of total peak number amount, peak area, peak height etc. grasp the ability and the mass spectral quality of protein and polypeptide in the mass spectrum of the protein polypeptide that magnetic bead kit is extracted.Ambient condition is suitable, and in the protein polypeptide mass spectrum of extraction, the peak number order can be many more, represents magnetic bead can filter out more protein from sample, provides bigger possibility for seeking differential protein; Peak area is more little, represents the tolerance range of protein peak high more, and the accuracy of later stage searching differential protein and proteinic evaluation just may be high more; The analysis that combines with signal to noise ratio of overall peak heights, noise cancellation signal is low, and the peak heights height shows that the mass spectrum quality is good, and the result is credible relatively.The 2nd, the ability of consistence, searching difference in the mass spectrum group.
Utilize the protein polypeptide that extracts to seek the differential protein relevant among the present invention of embodiment 5:SPE-C test kit, set up the process of more stable diagnostic model with disease.
SPE-C test kit among the present invention has very excellent performance, and it can extract abundant protein and polypeptide from serum, and sepn process is simple, and operation is convenient.Utilize SPE-C magnetic bead kit of the present invention, can make the mass spectroscopy of sample reach high-throughput, highly sensitive better plays a role biological mass spectrometry in clinical proteomics research.
Detailed process is as follows:
Software obtains data, handles, and obtains the mass spectrum of each sample; Software analysis obtains differential protein, sets up diagnostic model.Blind method is carried out sample process, detection; Software analysis and statistics determine its ownership disease group or normal group; Compare with clinical diagnosis; According to statistics, determine the blind property testing accuracy of unknown sample.The differential protein second order ms is identified.
Claims (10)
1.SPE-C system comprises the level pad, binding buffer liquid, cleaning buffer solution, the elutriant that are applicable to the SPE-C magnetic bead, wherein elutriant is the TFA of 0.1%-3%, and level pad, binding buffer liquid and cleaning buffer solution group are selected from:
0.001-2M pH is the Tris-HCL buffered soln of 7.0-9.0; Or
0.001-2M pH is the carbonate buffer solution of 9.0-11.0; Or
0.001-2M pH is the acetate buffer solution of 2.0-6.0; Or
0.001-2M pH is the glycine buffer of 2.0-6.0; Or
0.001-2M pH is the O-phthalic acid buffer of 2.0-6.0; Or
0.001-2M pH is the citrate buffer solution of 2.0-7.0; Or
0.001-2M pH is the TBS buffered soln of 7.0-9.0; Or
0.001-2M pH is the TE buffered soln of 2.0-7.0; Or
0.001-2M pH is the STE buffered soln of 7.0-9.0; Or
0.001-2M pH is the PBS buffered soln of 2.0-8.0; Or
0.001-2M pH is the SSC buffered soln of 7.0-9.0; Or
0.001-2M pH is the SSPE buffered soln of 7.0-9.0.
2. the described SPE-C of claim 1 system, wherein level pad is selected from: 0.25M, the Citrate trianion of pH2-6, PBS, acetate, glycine, phthalic acid or TE buffered soln;
Binding buffer liquid is selected from: 0.15M, the Citrate trianion of pH2-6, PBS, acetate, glycine, phthalic acid or TE buffered soln.
Cleaning buffer solution is selected from: 0.35M, the Citrate trianion of pH2-6, PBS, acetate, glycine, phthalic acid or TE buffered soln.
3. one kind by the prepared SPE-C test kit that is used for the separation of serum protein polypeptide of the SPE-C system of claim 1 or 2.
4. the described SPE-C test kit of claim 3 comprises the SPE-C magnetic bead that is used in conjunction with the separation of serum protein polypeptide.
5. the described SPE-C test kit of claim 4, wherein magnetic bead is the SPE-C magnetic bead.
6. the SPE-C test kit of the SPE-C system of claim 1 to 2, or claim 3 to 5 is used for the purposes of separation of serum protein polypeptide.
7. the purposes of claim 6 comprises that step is as follows:
(1) with SPE-C magnetic bead Balance Treatment, magnetic resolution is abandoned supernatant;
(2) the SPE-C magnetic bead combines with albumen, polypeptide generation specificity in the serum in binding buffer liquid, and magnetic resolution is abandoned supernatant;
(3) add cleaning buffered soln and clean magnetic bead, remove the anisogamy impurity of dtex;
(4) add elute soln, specific albumen and polypeptide in the wash-out serum, and measure peptide concentration;
(5) elutriant can be used for directly carrying out mass spectroscopy or frozenly carry out mass spectroscopy within-20 ℃, 24 hours.
8. the purposes of claim 7, wherein said purposes is to be used to seek the differential protein relevant with disease, sets up more stable diagnostic model.
9. the SPE-C test kit of the SPE-C system of claim 1 to 2, or claim 3 to 5 is used for the separation of serum protein polypeptide and according to the protein polypeptide mass spectrum that is separated to, screen significant differential protein carry out medical diagnosis on disease purposes.
10. the purposes of claim 9, protein polypeptide in the wherein said separation of serum, utilize the SPE-C test kit among the present invention, adopt MALDI-AUTO FLEX II TOF/TOF instrument, the interior consistence of protein polypeptide mass spectrum basic parameter, group that collects is analyzed.Obtain all proteic mass spectrums, by the software mass spectrum difference of patient or normal healthy controls relatively, obtain the otherness mass spectrum expression pattern of the two, the attribute that is used further to analyze unknown sample carries out the diagnosis of disease.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101936948A (en) * | 2010-08-06 | 2011-01-05 | 浙江省肿瘤医院 | Mass spectrum detection method of serum polypeptide |
| CN102830209A (en) * | 2012-09-11 | 2012-12-19 | 浙江大学 | Method for screening antidiabetic active compounds |
| CN111044731A (en) * | 2019-12-19 | 2020-04-21 | 北京科技大学 | Method for separating and enriching peptide impurities in polypeptide medicament by pulse incubation immunoreaction |
| CN112067388A (en) * | 2020-09-07 | 2020-12-11 | 苏州英芮诚生化科技有限公司 | High-throughput detection pretreatment method and kit for fat-soluble vitamins in blood plasma |
| CN114371065A (en) * | 2021-12-28 | 2022-04-19 | 上海固容生物科技有限公司 | Method for processing liquid biopsy sample of biological sample (magnetic bead separation method) and application thereof |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102331453A (en) * | 2011-04-15 | 2012-01-25 | 毅新兴业（北京）科技有限公司 | Method for detecting polypeptide in body fluid |
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2008
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101936948A (en) * | 2010-08-06 | 2011-01-05 | 浙江省肿瘤医院 | Mass spectrum detection method of serum polypeptide |
| CN102830209A (en) * | 2012-09-11 | 2012-12-19 | 浙江大学 | Method for screening antidiabetic active compounds |
| CN102830209B (en) * | 2012-09-11 | 2014-10-08 | 浙江大学 | Method for screening antidiabetic active compounds |
| CN111044731A (en) * | 2019-12-19 | 2020-04-21 | 北京科技大学 | Method for separating and enriching peptide impurities in polypeptide medicament by pulse incubation immunoreaction |
| CN112067388A (en) * | 2020-09-07 | 2020-12-11 | 苏州英芮诚生化科技有限公司 | High-throughput detection pretreatment method and kit for fat-soluble vitamins in blood plasma |
| CN114371065A (en) * | 2021-12-28 | 2022-04-19 | 上海固容生物科技有限公司 | Method for processing liquid biopsy sample of biological sample (magnetic bead separation method) and application thereof |
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