CN101907601A - Mass spectrum kit for hypertensive pregnancy biochemical marker and preparation method - Google Patents
Mass spectrum kit for hypertensive pregnancy biochemical marker and preparation method Download PDFInfo
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Abstract
The invention relates to a mass spectrum kit for a hypertensive pregnancy biochemical marker and a preparation method, and belongs to the technical field of protein detection. The kit consists of magnetic beads, adhesive liquid, cleaning solution, eluent, stabilizer and the like; and polypeptide and protein are enriched and purified from a complex biological sample. The kit comprises a small tube of adhesive liquid, a small tube of magnetic beads, a small tube of cleaning solution, a small tube of eluent and a small tube of stabilizer of energy-absorbing molecular saturated solution, wherein the small tubes are placed in a 4 DEG C refrigeration box. The mass spectrum kit can be used for in vitro sample detection and prognosis judgment. The characteristic protein peaks of the kit for judging the hypertensive pregnancy are 2,771, 3,218, 3,887, 4,192, 4,210, 5,101 and 5,689Da. The method is accurate, convenient and quick.
Description
Technical field
The invention belongs to technical field of protein detection, be particularly related to the kit preparation method that severe hypertension of pregnancy biological sample mesophytization label is analyzed of being used for, the novel biochemical label from complex biological sample in the enrichment and the purifying severe hypertension of pregnancy based on mass-spectrometric technique.
Background technology
No matter the normal function or the pathology characteristic that are cell all are somewhat dependent upon the expressed protein function of cell.Therefore, the difference of the protein of surveyor's expression in vivo can be used for diagnosis of vitro disease sample and examination, and finally is used for drug development and disease treatment.And to carry out the differentiation analysis of protein expression and function, requirement can reach the degree of differentiating the complex mixture of molecule in the cell.But many materials often exist with trace in the cell, and the method that is used for analyzing proteins at present has limitation at above-mentioned everyway, is difficult to carry out identification and analysis with these conventional meanses.Can overcome this technical disadvantages with the mass spectrum associating.
Hypertension of pregnancy disease gestation is distinctive, shows as the disease syndrome that occurs each the organ function disorder of elevation of blood pressure, urine protein and whole body after pregnant 20 weeks.The incidence of disease of serious preeclampsia: 3-5% is one of principal disease that causes the bad pregnancy outcome of female youngster perinatal period.Recent studies on method in recent years comprises: protein science, genomics, iipidomic, mRNA microarray technology etc., from the molecules level, the pathogenesis of further investigation hypertension of pregnancy disease, be expected to find the special biochemical indicator of hypertension of pregnancy disease, become the foundation of early diagnosis, the monitoring state of an illness and judgement disease prognosis.Proteomics is composition and the action rule of research genome at all protein of different time and space encoding, on histiocytic whole protein level, disclose the basic law of vital movement, obtain comprehensive and deep understanding of pair cell physiology, pathologic process and regulated and control network.The technology platform of using mainly contains two-dimensional gel electrophoresis isolation technics and analytical technique of mass spectrum.The former is mainly used in molecular weight 10-100kD Separation of Proteins, is difficult to detect for the protein of hydrophobicity, low abundance, low content.And mass spectrometer can be realized the following micromolecule of 10kD, low content, especially the isolation identification of the micro-regulation protein in the histocyte.The protein fingerprint pattern technology is a kind of new Separation of Proteins, authentication method.Its principle is: utilize the laser pulse radiation to make the analyte desorption in the protein-chip pond form charged ion, the time that the ion of different mass-to-charge ratioes flies in the instrument field is different in size, after software processes, draws out the protein spectrogram accurately.Its advantage is: can directly from primary sample, catch, detects and analyze, and highly sensitive, sensing range wide (500～500000), required sample volume little (0.5～400 μ L); Detect easyly, whole mensuration process is generally 30min, and susceptibility and specificity height; Simultaneously can not destroy the protein of being measured; Reliable results is repeatable high.This technology not only can filter out the special biomarker with disease association, also can find the protein fingerprint pattern with the combination of the different modes of disease association, for clinical early diagnosis, the discovery of medicine target of disease, curative effect is judged and prognosis provides foundation.
Carrying out not finding also when proteomic image is analyzed that is used for the standardization mass spectrometry kit that severe hypertension of pregnancy biochemical marker detects at present.
Summary of the invention
The objective of the invention is to overcome the weak point of prior art, a kind of mass spectrometry kit and preparation method who is used to detect clinical severe hypertension of pregnancy biochemical marker proposed, this kit provides new approach for the early detection of the severe hypertension of pregnancy, and for finding that further new severe hypertension of pregnancy biological marker provides the foundation.
The mass spectrometry kit that is used to detect severe hypertension of pregnancy biological marker that the present invention proposes is characterized in that this kit comprises:
One tubule slurry, 10～30 μ l;
One tubule mass spectrum standardization quality controlled serum (slurry), 10～30 μ l;
One tubule MB-WCX magnetic bead or the affine surface of metal (MB-IMAC Fe) magnetic bead 30～50 μ l;
One tubule cleaning fluid, 300～500 μ l, this cleaning fluid is 50～100mM PBS or NaAc, the pH value is 4.0～5.0;
[cleaning fluid pH value is 3.0～6.0 o'clock, and protein fingerprint detects finds that protein fingerprint detected best (protein peak is maximum) when the pH value was 4.0～5.0 cleaning fluids].
One tubule eluant, 10～50 μ l, this eluant is the aqueous solution of 1～5% trifluoroacetic acid;
One tubule stabilizing agent, 5～10 μ l, this stabilizing agent is made of in the aqueous solution that contains 30～60% acetonitriles and 0.5～1% trifluoroacetic acid the energy absorption molecular melting, and this energy absorption molecule can adopt any among cinnamic acid derivative, sinapic acid, the dihydroxy-benzoic acid; Above-mentioned each tubule places 4 ℃ of refrigerating box.
The above-mentioned reagents for mass spectrogram box preparation method that the present invention proposes is characterized in that this method may further comprise the steps:
1) contains the preparation of the mass spectrum standardization quality controlled serum (slurry) of slurry: the O type serum (slurry) of the male sex, women's equivalent is diluted in makes mass spectral standardization quality controlled serum (slurry) in the slurry, add 10% bovine insulin (bovine insulin, 5733Da), and the serum that will dilute (slurry) be distributed in 10～30 μ l tubules; Wherein, serum (slurry): slurry=1: 2, described slurry are 9M Urea, 2%CHAPS, and 50mM Tris-HCL, pH 7.0～9.0;
50～100mM PBS that 2) will buy (Phosphate-Buffered Saline) or NaAc, the pH value is that the eluant of 4.0～5.0 cleaning fluids, 1～5% trifluoroacetic acid is distributed into 300～500 μ l tubules and 10～50 μ l tubules respectively;
3) make stabilizing agent in the aqueous solution with energy absorption molecular melting 30～60% acetonitriles and 0.5～1% trifluoroacetic acid, and being distributed into 5～10 μ l tubules, this energy absorption molecule can adopt any among cinnamic acid derivative, sinapic acid, the dihydroxy-benzoic acid;
4) each tubule that above-mentioned branch is installed places 4 ℃ of congealeres.
The present invention also proposes the application that described kit is judged the prognosis of severe hypertension of pregnancy biological marker.
Described kit is 2771,3218,3887,4192,4210,5101 and 5689Da (Fig. 1) according to the several characteristic protein peak, the susceptibility 100% of double-blind severe hypertension of pregnancy biological marker sample, and specificity is 100%.
The application experiment step that mass spectrometry kit detects severe hypertension of pregnancy biological marker comprises:
1. biological sample is diluted 30～50 times with slurry earlier, with the abundant mixing of sample;
2. above-mentioned sample 100 μ l are added in the PCR pipe that installs magnetic bead, put on the magnetic processor, hatched 20～40 minutes, and removed liquid for 15～25 ℃;
3. add 100 μ l cleaning fluids to the PCR pipe that installs magnetic bead, put and hatch 1～5 minute on the magnetic processor, remove liquid, repeat twice of aforesaid operations;
4. add 10 μ l eluants 1～5 minute, the wash-out sample is to supernatant;
5. get 5 μ l supernatants and move in another PCR pipe, add the abundant mixing of the 5 saturated stabilizing agents of μ l energy absorption molecule;
6. get 1 μ l mixed solution application of sample to mass spectrum special-purpose metal sheet (the 3x3mm circular hole is arranged), the air dry sheet metal;
7. above-mentioned sheet metal is added in the mass spectrometer, will generate mass spectrum;
8. outside use peptide molecule quality standard is come the correction mass accuracy, and all sample standard deviations carry out double and detect to reduce experimental error.
Above-mentioned biological marker utilizes a mass spectrometer to detect.The exactness high in quality of this equipment is about+and/-1%.
Severe hypertension of pregnancy biological marker at first can be had magnetic bead (the Magnetic Bead that can combine with severe hypertension of pregnancy biological marker, be called for short MB, as names of an article such as MB-WCX, MB-IMAC Fe) absorption surface catches, non-adsorbate can be from wash-out on the magnetic bead, and the severe hypertension of pregnancy biological marker that is adsorbed onto the magnetic bead base is detected in mass spectrometer.The source takes place by ion in severe hypertension of pregnancy biological marker, as laser, is ionized, and the ion of generation is experienced the collector collection by an ion, and mass analyzer is analyzed those ions that passes through then.Afterwards, detecting device is a mass-to-charge ratio with the ion information translation that detects.Quantitatively property control and mass spectrum laser energy regulation and control: before each test, with mass spectral standardization quality controlled serum, will be used for quantitative standards peak bovine insulin in the standardization quality controlled serum, 5733Da intensity transfers to the maximal value of 50% mass signal intensity.The detection of severe hypertension of pregnancy biological marker is significantly with relevant with the detection of signal intensity.Like this, the quantity of severe hypertension of pregnancy biological marker and quality can be detected.
Mass spectrum generates time of flight spectrum to the analysis of analysans.The independent pulse signal that sample of ionization energy attack produces is not represented in the final analysis of this time of flight spectrum, but the signal sum of a series of pulses.Reduce interference like this, and increased dynamic range.These flight time data are subjected to the influence of data processing software.Data processing mainly comprises conversion flight time and mass-to-charge ratio and produces mass spectrum in the software, reduces baseline and reduces the side-play amount of instrument and filtration high frequency noise and alleviate high frequency noise.
By being surveyed the data that produce, the health check-up of severe hypertension of pregnancy biological marker can utilize the DAP of computing machine to analyze.These data of this computer program analysis to be showing the quantity of detected severe hypertension of pregnancy biological marker, and the intensity of shows signal and determine the molecular weight of each detected severe hypertension of pregnancy biological marker.Data analysis can also comprise the signal intensity of a series of definite severe hypertension of pregnancy biological marker and correct data departing from predetermined statistical distribution state.For example, by the height of calculating with each peak value of some parameter correlation, but the peak that standard observes.This parameter may be the unessential interference that is produced by chemical constitutions such as instrument and similar energy absorption molecules, and this can be provided with zeroing.
Computing machine can convert calculation result data to various forms and show.Its standard spectrum can represent, but has only peak height and quality information to keep in bands of a spectrum in one form, produces a figure more clearly, and makes and have easier the manifesting of severe hypertension of pregnancy biological marker of molecular weight much at one.In another form, two or more spectrums relatively are convenient to highlight unique severe hypertension of pregnancy biological marker and are higher or lower than the severe hypertension of pregnancy biological marker of calibration sample with those.
Analysis generally comprises the evaluation at peak the collection of illustrative plates of the signal that displaying obtains from analysans.The peak can be selected by view, and software is available, and it is detected peaks automatically.Generally speaking, this software is tested and appraised signal and has signal to noise ratio (S/N ratio) and be higher than one and select threshold value and mark in the such mode of quality at the peak at the barycenter place of peak-to-peak signal to operate.In an effective program, more many spectral lines appear in the mass spectrum some same in a certain selected scope peaks with identification.A version of this software is assembled all peaks that appear at each the bar spectrum in definite mass range, near all peaks quality of appointment (mass-to-charge ratio) quality (mass-to-charge ratio) intermediate value bunch.
The severe hypertension of pregnancy biological marker that uses in the invention is that magnetic bead is caught.These severe hypertension of pregnancy biological markers are further to measure the identity that its different molecular weight knows that they are specific by mass spectrum (mass spectrometry).
Severe hypertension of pregnancy biological marker detects the polypeptide protein screening.By the haemocyanin fingerprint peaks that Mann-Whitney U check analysis hundreds of kind magnetic bead (MB-WCX, MB-IMAC Fe) is caught, find that following protein fingerprint or mass-spectrogram can be used to distinguish the serum of normal and severe hypertension of pregnancy biological marker.Peak C V＞30% or p＜0.05 is for there being significant difference:
The table one severe hypertension of pregnancy (pregnant height) changes with normal serum protein lotus
Therefrom filter out the several characteristic protein peak, 2771,3218,3887,4192,4210,5101 and the testing model formed of 5689Da difference peak severe hypertension of pregnancy patient and the blind screening of healthy population are tested, analyze the mass spectrum result of 50 parts of severe hypertension of pregnancy samples with this classification, wherein distinguish correct for 50 parts, 0 part of error differentiating, susceptibility are 100%; Distinguish correctly for 100 parts in 100 parts of control samples, 0 part of error differentiating, specificity is 100%.
Utilize the experimental result of the affine surface of metal (MB-IMAC Fe) magnetic bead or chip consistent with the experimental result of negative ion magnetic bead (MB-WCX) or chip.
The present invention can be used for the quantitative control of the external severe hypertension of pregnancy biological marker Mass Spectrometer Method method of cell in vitro and Noninvasive, is used for the severe hypertension of pregnancy biological marker detection method of mass spectrometry clinical as the severe hypertension of pregnancy biological marker kit of the body fluid that exsomatizes.Can detect a plurality of severe hypertension of pregnancy biomarker protein matter biological indication marks groups and mass spectrum polypeptide collection of illustrative plates.
The external detection method of kit among the present invention and method and other Noninvasives relatively has following characteristics:
(1) accurate
Mass spectrum is directly analyzed very strong accuracy.Because protein is made up of amino acid, and amino acid whose average quality is known.Mass-to-charge ratio (m/z) is that 2771,3218,3887,4192,4210,5101 and 5689 albumen is expressed in the patient that the severe hypertension of pregnancy takes place and obviously increased, for the good predictability of having of the severe hypertension of pregnancy.
(2) convenient
The sorbent used holder of the method for supported matrix is a magnetic bead.Separate magnetic bead and sample with magnetic separator, need not centrifugal sample.
(3) quick
When detecting, need not protein is checked order with protein fingerprint method provided by the invention.
Description of drawings
The representative protein fingerprint peak that raises in Fig. 1 severe hypertension of pregnancy serum
Embodiment
The present invention will be described further in conjunction with specific embodiments, and these examples only are used for illustration purpose, and are not used in the restriction scope of the invention.Embodiment 1
The mass spectrometry kit embodiment 1 of the severe hypertension of pregnancy biological marker that the present invention proposes comprises:
One tubule slurry, 10～30 μ l;
One tubule mass spectrum standardization quality controlled serum (slurry), 10～30 μ l;
One tubule MB-WCX magnetic bead or the affine surface of metal (MB-IMAC Fe) magnetic bead (Beijing Sai Erdi company product) 30～50 μ l;
One tubule cleaning fluid, 300～500 μ l, this cleaning fluid is 50～100mM PBS or NaAc, the pH value is 4.0～5.0;
One tubule eluant, 10～50 μ l, this eluant is the aqueous solution of 1～5% trifluoroacetic acid;
One tubule stabilizing agent, 5～10 μ l, this stabilizing agent is made of in the aqueous solution that contains 30～60% acetonitriles and 0.5～1% trifluoroacetic acid the energy absorption molecular melting, and this energy absorption molecule can adopt any among cinnamic acid derivative, sinapic acid, the dihydroxy-benzoic acid; Above-mentioned each tubule places 4 ℃ of refrigerating box.
The above-mentioned reagents for mass spectrogram box preparation method that the present invention proposes is characterized in that this method may further comprise the steps:
1) contains the preparation of the mass spectrum standardization quality controlled serum (slurry) of slurry: the O type serum (slurry) of the male sex, women's equivalent is diluted in makes mass spectral standardization quality controlled serum (slurry) in the slurry, add 10% bovine insulin (bovine insulin, 5733Da), and the serum that will dilute (slurry) be distributed in 10～30 μ l tubules; Wherein, serum (slurry): slurry=1: 2, described slurry are 9M Urea, 2%CHAPS, and 50mM Tris-HCL, pH 7.0～9.0;
50～100mM PBS that 2) will buy (Phosphate-Buffered Sal ine) or NaAc, the pH value is that the eluant of 4.0～5.0 cleaning fluids, 1～5% trifluoroacetic acid is distributed into 300～500 μ l tubules and 10～50 μ l tubules respectively;
3) make stabilizing agent in the aqueous solution with energy absorption molecular melting 30～60% acetonitriles and 0.5～1% trifluoroacetic acid, and being distributed into 5～10 μ l tubules, this energy absorption molecule can adopt any among cinnamic acid derivative, sinapic acid, the dihydroxy-benzoic acid;
4) each tubule that above-mentioned branch is installed places 4 ℃ of congealeres.
The present invention is used for severe hypertension of pregnancy biological marker and detects the polypeptide protein screening experiment
(1) experimental technique
The application experiment step that mass spectrometry kit detects severe hypertension of pregnancy biological marker comprises:
1. biological sample is diluted 30～50 times with slurry earlier, with the abundant mixing of sample;
2. above-mentioned sample 100 μ l are added in the PCR pipe that installs magnetic bead, put on the magnetic processor, hatched 20～40 minutes, and removed liquid for 15～25 ℃;
3. add 100 μ l cleaning fluids to the PCR pipe that installs magnetic bead, put and hatch 1～5 minute on the magnetic processor, remove liquid, repeat twice of aforesaid operations;
4. add 10 μ l eluants 1～5 minute, the wash-out sample is to supernatant;
5. get 5 μ l supernatants and move in another PCR pipe, add the abundant mixing of the 5 saturated stabilizing agents of μ l energy absorption molecule;
6. get 1 μ l mixed solution application of sample to mass spectrum special-purpose metal sheet (the 3x3mm circular hole is arranged), the air dry sheet metal;
7. above-mentioned sheet metal is added in the mass spectrometer, will generate mass spectrum;
8. outside use peptide molecule quality standard is come the correction mass accuracy, and all sample standard deviations carry out double and detect to reduce experimental error.
The table one severe hypertension of pregnancy (pregnant height) changes with normal serum protein lotus
Data aggregation
The Au sheet of handling well is inserted MALDI-TOF-MS carry out the protein spectrum analysis.Before reading of data, proofread and correct mass spectrometer with the chip that is added with the all-in-one standard protein, make molecular weight error＜0.1%.The major parameter of reading apparatus is set in this research, and the highest detection molecular weight is 50kDa, optimizes molecular weight ranges 1000～15000Da, optimal accumulated center 8000Da, and data acquisition parameters scope 20～80, collection adds up to 130 times.Set in the software and read the sheet program, with reading of data.Protein fingerprint spectrum with in the correction of the interior mark peak in the serum raw data makes molecular weight error＜0.01%, thus the accurate protein fingerprint spectrum that obtains.
Statistical analysis
All peak spectrums of application software analyzing and processing form protein fingerprint spectrum.All collection of illustrative plates have all carried out standardization, and are unified to they own whole population of ions (summation of peak area).To be used for the maximal value that the quantitative standards peak intensity transfers to 40～50% signal intensities in the standardization quality controlled serum, and proofread and correct, then carried out again in " minimizing baseline ", definition protein peak (s/n＞5, minimum peak intensity＞1.6) with the most significant peak.Analyze the protein peak between all 1～50kDa, and scrutiny each corresponding peak, calculate average, standard error (SD) and the coefficient of variation (CV%) at peak.Check the relatively protein peak of paired sample with Mann-Whitney U, calculate the p value.
Severe hypertension of pregnancy biological marker detects the polypeptide protein screening.By the haemocyanin fingerprint peaks of Mann-Whitney U check analysis hundreds of kind magnetic capture, find that following protein fingerprint or mass-spectrogram can be used to distinguish the serum of normal and severe hypertension of pregnancy biological marker.Peak C V＞30% or p＜0.05 is for there being significant difference (table one).
Therefrom filter out the several characteristic protein peak, 2771,3218,3887,4192,4210,5101 and the testing model formed of 5689Da difference peak severe hypertension of pregnancy patient and the blind screening of healthy population are tested, analyze the mass spectrum result of 50 parts of severe hypertension of pregnancy samples with this classification, wherein distinguish correct for 50 parts, 0 part of error differentiating, susceptibility are 100%; Distinguish correctly for 100 parts in 100 parts of control samples, 0 part of error differentiating, specificity is 100%.
Utilize the experimental result of the affine surface of metal (MB-IMAC Fe) magnetic bead or chip consistent with the experimental result of negative ion magnetic bead (MB-WCX) or chip.
List of references:
[1]National?High?Blood?Pressure?Education?Program?Working?Group.Am.J?Obstet?Gynecol.2000，183：S1-S2.
[2]Baumwell?S，Karumanchi?SA，Pre-eclampsia：clinical?manifestations?and?molecular?mechanisms.Nephron?Clin?Pract.2007.106：c72-c81.
[3] Xu Yang. the progress [J] of protein fingerprint pattern technology in laboratory diagnosis and clinical medicine. preclinical medicine and clinical, 2007,27 (2): 134-142.
[4]Hayman?R.A，Johnson?W.I，Baker?P.The?preliminary?characterization?of?a?vasoactive?circulating?factor(s)in?preeclampsia.American?Journal?of?Obstetrics?and?Gynecology.2001，184：1196-1203.
[5]Jenny?M，Maureen?M，B.REED，N.Harris，et?al.Use?of?proteomic?patterns?as?a?novel?screening?tool?in?pre-eclampsia.Journal?of?Obstetrics?and?Gynaecology.2004，24(8)：873-874.
[6]Li-zhou?Sun，Na-na?Yang，Wei?De，Yun-shan?Xiao.Proteomic?Analysis?of?Proteins?Differentially?Expressed?in?Preeclamptic?Trophoblasts.Gynecol?Obstet?Invest.2007，64：17-23.
After having read foregoing of the present invention, those skilled in the art can make various changes or modifications the present invention, and these equivalent form of values should fall within the application's appended claims institute restricted portion equally.
Claims (6)
1. a mass spectrometry kit that detects severe gestation hypertension biochemical marker is characterized in that, this kit comprises:
One tubule slurry, 10～30 μ l;
One tubule mass spectrum standardization quality controlled serum (slurry), 10～30 μ l;
One tubule MB-WCX magnetic bead or the affine surface of metal (MB-IMAC Fe) magnetic bead 30～50 μ l;
One tubule cleaning fluid, 300～500 μ l are 50～100mM PBS or NaAc, and the pH optimal value is 4.0～5.0;
One tubule eluant, 10～50 μ l, this eluant is the aqueous solution of 1～5% trifluoroacetic acid;
One tubule stabilizing agent, 5～10 μ l, this stabilizing agent is made of in the aqueous solution that contains 30～60% acetonitriles and 0.5～1% trifluoroacetic acid the energy absorption molecular melting, and this energy absorption molecule can adopt any among cinnamic acid derivative, sinapic acid, the dihydroxy-benzoic acid; Above-mentioned each tubule places 4 ℃ of refrigerating box.
2. kit as claimed in claim 1 is characterized in that, any among described energy absorption molecule employing cinnamic acid derivative, sinapic acid, the dihydroxy-benzoic acid.
3. a method for preparing mass spectrometry kit as claimed in claim 1 is characterized in that, this method may further comprise the steps:
1) contains the preparation of the mass spectrum standardization quality controlled serum (slurry) of slurry: the O type serum (slurry) of the male sex, women's equivalent is diluted in makes mass spectral standardization quality controlled serum (slurry) in the slurry, add 10% bovine insulin (bovine insulin, 5733Da), and the serum that will dilute (slurry) be distributed in 10～30 μ l tubules; Wherein, serum (slurry): slurry=1: 2, described slurry are 9M Urea, 2%CHAPS, and 50mM Tris-HCL, pH 7.0～9.0;
50～100mM PBS that 2) will buy (Phosphate-Buffered Saline) or NaAc, the pH value is that the eluant of 4.0～5.0 cleaning fluids, 1～5% trifluoroacetic acid is distributed into 300～500 μ l tubules and 10～50 μ l tubules respectively;
3) make stabilizing agent in the aqueous solution with energy absorption molecular melting 30～60% acetonitriles and 0.5～1% trifluoroacetic acid, and being distributed into 5～10 μ l tubules, this energy absorption molecule can adopt any among cinnamic acid derivative, sinapic acid, the dihydroxy-benzoic acid;
4) each tubule that above-mentioned branch is installed places 4 ℃ of congealeres.
4. method as claimed in claim 3 is characterized in that, any among described energy absorption molecule employing cinnamic acid derivative, sinapic acid, the dihydroxy-benzoic acid.
5. application of wanting kit as described in 1 that the severe gestation hypertension is judged as right.
6. application as claimed in claim 5, described kit is judged the severe gestation hypertension; This kit is 2771,3218,3887,4192,4210,5101 and 5689Da according to the several characteristic protein peak.
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Cited By (3)
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| CN102375063A (en) * | 2010-08-23 | 2012-03-14 | 湖州赛尔迪生物医药科技有限公司 | Immune mass spectrometric kit of common proteins and preparation method thereof |
| CN104597114A (en) * | 2015-01-21 | 2015-05-06 | 华中师范大学 | Mass correction kit and mass correction method for low-mass area of high-resolution mass spectrometer in negative ion mode |
| CN105067821A (en) * | 2015-08-10 | 2015-11-18 | 国家纳米科学中心 | Method for realizing target fishing and characterization through small molecule microarray |
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| CN101153872A (en) * | 2006-09-29 | 2008-04-02 | 许洋 | Novel reagent kit for detecting and estimating critical patients and method thereof |
| CN101303360A (en) * | 2008-04-25 | 2008-11-12 | 许洋 | Mass spectrogram antibody kit of human chorionic gonadotrophin isomer and preparation method thereof |
| EP2073011A1 (en) * | 2007-12-20 | 2009-06-24 | Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. | Means and methods of processing biological samples |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101153872A (en) * | 2006-09-29 | 2008-04-02 | 许洋 | Novel reagent kit for detecting and estimating critical patients and method thereof |
| EP2073011A1 (en) * | 2007-12-20 | 2009-06-24 | Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. | Means and methods of processing biological samples |
| CN101303360A (en) * | 2008-04-25 | 2008-11-12 | 许洋 | Mass spectrogram antibody kit of human chorionic gonadotrophin isomer and preparation method thereof |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102375063A (en) * | 2010-08-23 | 2012-03-14 | 湖州赛尔迪生物医药科技有限公司 | Immune mass spectrometric kit of common proteins and preparation method thereof |
| CN104597114A (en) * | 2015-01-21 | 2015-05-06 | 华中师范大学 | Mass correction kit and mass correction method for low-mass area of high-resolution mass spectrometer in negative ion mode |
| CN104597114B (en) * | 2015-01-21 | 2016-03-30 | 华中师范大学 | The mass calibration kit of high-resolution mass spectrometer negative ion mode low mass region and bearing calibration |
| CN105067821A (en) * | 2015-08-10 | 2015-11-18 | 国家纳米科学中心 | Method for realizing target fishing and characterization through small molecule microarray |
| CN105067821B (en) * | 2015-08-10 | 2017-02-22 | 国家纳米科学中心 | Method for realizing target fishing and characterization through small molecule microarray |
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Application publication date: 20101208 |