CN1657941A - Use of investigating multiple antigen label combined by antibody absorption surface material and mass spectrum - Google Patents
Use of investigating multiple antigen label combined by antibody absorption surface material and mass spectrum Download PDFInfo
- Publication number
- CN1657941A CN1657941A CN 200410004403 CN200410004403A CN1657941A CN 1657941 A CN1657941 A CN 1657941A CN 200410004403 CN200410004403 CN 200410004403 CN 200410004403 A CN200410004403 A CN 200410004403A CN 1657941 A CN1657941 A CN 1657941A
- Authority
- CN
- China
- Prior art keywords
- antibody
- absorption surface
- protein
- mark
- surface material
- 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
- 102000036639 antigens Human genes 0.000 title claims abstract description 25
- 108091007433 antigens Proteins 0.000 title claims abstract description 25
- 239000000427 antigen Substances 0.000 title claims abstract description 24
- 239000000463 material Substances 0.000 title claims description 22
- 238000010521 absorption reaction Methods 0.000 title claims description 20
- 238000001819 mass spectrum Methods 0.000 title abstract description 13
- 201000010099 disease Diseases 0.000 claims abstract description 22
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 22
- 238000000034 method Methods 0.000 claims abstract description 15
- 239000012472 biological sample Substances 0.000 claims abstract description 5
- 239000000090 biomarker Substances 0.000 claims description 38
- 102000004169 proteins and genes Human genes 0.000 claims description 32
- 108090000623 proteins and genes Proteins 0.000 claims description 32
- 230000000890 antigenic effect Effects 0.000 claims description 15
- 239000000523 sample Substances 0.000 claims description 14
- 238000004458 analytical method Methods 0.000 claims description 12
- 238000001514 detection method Methods 0.000 claims description 8
- 239000008280 blood Substances 0.000 claims description 4
- 238000004949 mass spectrometry Methods 0.000 claims description 4
- 238000002798 spectrophotometry method Methods 0.000 claims description 4
- 210000004369 blood Anatomy 0.000 claims description 3
- 210000001124 body fluid Anatomy 0.000 claims 2
- 239000010839 body fluid Substances 0.000 claims 2
- 210000000056 organ Anatomy 0.000 claims 2
- 230000028327 secretion Effects 0.000 claims 2
- 101710153593 Albumin A Proteins 0.000 claims 1
- 206010057248 Cell death Diseases 0.000 claims 1
- 101710120037 Toxin CcdB Proteins 0.000 claims 1
- 239000012491 analyte Substances 0.000 claims 1
- 239000011324 bead Substances 0.000 claims 1
- 239000000919 ceramic Substances 0.000 claims 1
- 230000009089 cytolysis Effects 0.000 claims 1
- 239000011521 glass Substances 0.000 claims 1
- 239000006249 magnetic particle Substances 0.000 claims 1
- 239000002184 metal Substances 0.000 claims 1
- 229920000642 polymer Polymers 0.000 claims 1
- 238000001179 sorption measurement Methods 0.000 abstract description 4
- 238000005259 measurement Methods 0.000 abstract 1
- 235000018102 proteins Nutrition 0.000 description 22
- 210000002966 serum Anatomy 0.000 description 11
- 239000012634 fragment Substances 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- 150000001413 amino acids Chemical group 0.000 description 6
- 238000001228 spectrum Methods 0.000 description 6
- 208000006379 syphilis Diseases 0.000 description 6
- 210000001685 thyroid gland Anatomy 0.000 description 6
- 150000002500 ions Chemical class 0.000 description 5
- 102100023635 Alpha-fetoprotein Human genes 0.000 description 4
- 101710083889 Alpha-fetoprotein Proteins 0.000 description 4
- 102000004420 Creatine Kinase Human genes 0.000 description 4
- 108010042126 Creatine kinase Proteins 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- 108010044467 Isoenzymes Proteins 0.000 description 4
- 102000004357 Transferases Human genes 0.000 description 4
- 108090000992 Transferases Proteins 0.000 description 4
- 102000002070 Transferrins Human genes 0.000 description 4
- 108010015865 Transferrins Proteins 0.000 description 4
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 4
- 239000005556 hormone Substances 0.000 description 4
- 229940088597 hormone Drugs 0.000 description 4
- PCMORTLOPMLEFB-ONEGZZNKSA-N sinapic acid Chemical compound COC1=CC(\C=C\C(O)=O)=CC(OC)=C1O PCMORTLOPMLEFB-ONEGZZNKSA-N 0.000 description 4
- PCMORTLOPMLEFB-UHFFFAOYSA-N sinapinic acid Natural products COC1=CC(C=CC(O)=O)=CC(OC)=C1O PCMORTLOPMLEFB-UHFFFAOYSA-N 0.000 description 4
- 230000001646 thyrotropic effect Effects 0.000 description 4
- 101710088194 Dehydrogenase Proteins 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 3
- 102000008946 Fibrinogen Human genes 0.000 description 3
- 108010049003 Fibrinogen Proteins 0.000 description 3
- 206010019280 Heart failures Diseases 0.000 description 3
- 235000001014 amino acid Nutrition 0.000 description 3
- 230000029087 digestion Effects 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 229940012952 fibrinogen Drugs 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 230000035935 pregnancy Effects 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 108010051815 Glutamyl endopeptidase Proteins 0.000 description 2
- 101710159910 Movement protein Proteins 0.000 description 2
- 102100024147 Protein phosphatase 1 regulatory subunit 14A Human genes 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000013016 damping Methods 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- DAPOMFMFTBUAKL-UHFFFAOYSA-N 2-ethylpyridine-3-thiol Chemical compound CCC1=NC=CC=C1S DAPOMFMFTBUAKL-UHFFFAOYSA-N 0.000 description 1
- 102000009123 Fibrin Human genes 0.000 description 1
- 108010073385 Fibrin Proteins 0.000 description 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 1
- 108700020403 Human Immunodeficiency Virus Type 1 p24 Proteins 0.000 description 1
- 241000713772 Human immunodeficiency virus 1 Species 0.000 description 1
- 206010061598 Immunodeficiency Diseases 0.000 description 1
- 208000029462 Immunodeficiency disease Diseases 0.000 description 1
- 102000003855 L-lactate dehydrogenase Human genes 0.000 description 1
- 108700023483 L-lactate dehydrogenases Proteins 0.000 description 1
- 239000006096 absorbing agent Substances 0.000 description 1
- 239000002156 adsorbate Substances 0.000 description 1
- 230000036436 anti-hiv Effects 0.000 description 1
- 230000001014 anti-treponemal effect Effects 0.000 description 1
- 230000009830 antibody antigen interaction Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000012148 binding buffer Substances 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000004590 computer program Methods 0.000 description 1
- 230000009514 concussion Effects 0.000 description 1
- 238000012937 correction Methods 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 239000007857 degradation product Substances 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000002405 diagnostic procedure Methods 0.000 description 1
- 239000012470 diluted sample Substances 0.000 description 1
- 238000009509 drug development Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 229950003499 fibrin Drugs 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000002795 fluorescence method Methods 0.000 description 1
- 230000007813 immunodeficiency Effects 0.000 description 1
- 238000010166 immunofluorescence Methods 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000004853 protein function Effects 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 238000012113 quantitative test Methods 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Landscapes
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
This invention relates to a method which is through the antigen mark which is caught by the immune body adsorption surface matter on, and analyzes with the mass spectrum examines. Adsorbs the many kinds of antigen mark in on a many kinds of immune bodies adsorptions surface, and carries on the mass spectrum analysis to the capture antigen mark. May examine many antigen mark group. This method may use examining the antigen mark combination in the biological sample. Examines the measurement with these antigen mark combinations to be possible at the same time to distinguish the many kinds of diseases.This method is accurate, convenience also quickly.
Description
Technical field
The present invention relates to protein analysis method in a kind of new biological sample, a kind of biomarker of catching by antibody absorption surface material, and come the detection of biological mark with mass spectrophotometry.The present invention has been improved the immune analysis method means widely referred in this, and is more accurate than present routine biochemistry inspection (enzyme-linked method or fluorescence method).Say that more properly this invention relates to various ways biomarker (biomarkers) or antigenic mark, and these antigens or biomarker can be used to distinguish multiple disease marker is disposable with higher specificity and sensitivity.The present invention can be applied to clinical examination.
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, identify the difference of disease expressed protein in human body, can be used for medical diagnosis on disease and examination, and finally be 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 analysing protein at present has limitation at above-mentioned everyway, is difficult to multiple disease is carried out qualitative evaluation and quantitative test simultaneously with these conventional meanses.
As, hospital requires to detect specifically the known mark of multiple common disease.But the reagent for preparing multiple specificity incorporation of markings and can identify mark in the potpourri of complexity needs the plenty of time, and this has hindered the development of this type of diagnostic method.At present, also there is not a kind of method four kinds of disease markers can be detected simultaneously.
ELISA (enzyme-linked immunosorbent assay) kit utilizes antibody and the reaction of enzyme mark can be used to detect a kind of disease marker.Utilize antibody and three fluorescence, can accomplish to detect three disease markers at most, but three above disease markers just can't detect simultaneously.Utilize antibody absorption surface material and antibody and mass spectrometer use in conjunction, promptly can solve the disease antigen mark of differentiating simultaneously more than three kinds.Say for example, to resist HBV (HBsAg) antibody, HCV antigen/antibody combination, anti-HIV p24 antigen (Ribas SG et al.Performance of a quantitative humanimmunodeficiency virus type 1 p24 antigen assay on various HIV-1 subtypes for thefollow-up of human immunodeficiency type 1 seropositive individuals.J Virol Methods2003; 113:29-34) and anti-syphilis antibody (anti-treponemal 17 kDa protein) (George R et al.Ananalysis of the value of some antigen-antibody interactions used as diagnosticindicators in a treponemal Western blot (TWB) test for syphilis.J Clin Lab Immunol1998; Antibody combined being tagged on Protein A or the G antibody absorption surface material such as 50:27-44).Because the molecular weight of every kind of sick viral antigen of special body antibody capture is different, so when Protein A/G-antibody and mass spectrometer use in conjunction, mass spectrometer has just separated multiple antigen easily simultaneously.Reasoning thus, if select multiple specific antibody simultaneously, and the antigen molecular of these antibody institute combinations is different, then the present invention can distinguish countless versions disease antigen mark simultaneously.
Summary of the invention
The objective of the invention is to set up a kind of purposes that in biological sample, detects multiple disease biomarker or antigenic mark.This invention relates to biomarker (biomarkers), and these biomarkers can be distinguished multiple disease simultaneously by the higher specificity and the mass spectrum of sensitivity.
The present invention relates to a kind of mark specific antibody that passes through to antibody absorption surface material (Protein A and G or Mercapto-Ethyl-Pyridine), and detect multiple disease biomarker simultaneously with mass spectrophotometry.
Biomarker among the present invention is to utilize a mass spectrometer to do last evaluation.The exactness high in quality of this equipment is about+and/-0.1%.
Antibody absorption surface material: with Protein A and G antibody absorption surface material is representative (Protein A and G has capture antibody Fc section function).Protein A and G base and antibodies with absorbing agent function, biomarker/antigen protein in the antibodies serum.Through one section time enough, biomarker/antigen can be combined with antibody-ProteinA/G.The material that the base flush away does not adsorb.
Biomarker at first can be had the special antibody-Protein A and G absorption surface material that can combine with biomarker catches, and non-adsorbate can be from wash-out on the absorption surface material.Add reductive agent then, antigen is separated with antibody, the supernatant that will contain antigen is detected in flight mass spectrometer.The source takes place by ion in biomarker, 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.The detection of biomarker is significantly with relevant with the detection of signal intensity.Like this, the quantity of biomarker and quality can be detected.
Flight 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.
Can utilize the DAP of computing machine to analyze by the data that the detection to biomarker produces.These data of this computer program analysis to be showing the quantity of detected label, and the intensity of shows signal and determine the molecular weight of each detected biomarker.Data analysis can also comprise the signal intensity of a series of definite biomarker 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 value that standard is measured.This parameter may be the 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 computational data to various forms and show.Its standard spectrum can be expressed as two kinds: in a kind of form, have only peak height and quality information to keep in bands of a spectrum, produce one and scheme, and make more clearly to have easier the manifesting of biomarker of molecular weight much at one.In another form, two or more spectrums relatively are convenient to highlight the unique biological label and are higher or lower than the biomarker 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 multiple disease biomarker that uses in the invention is caught by multiple special antibody.These biomarkers are by measuring its different molecular weight by mass spectrum (mass spectrometry).
On the adsorption site that a sample need be placed on the detection of biomarker, then clean.On adsorption site, add SINAPINIC acid and allow its drying.Then, with mass spectroscopy it is analyzed, and a legacy figure who has shown protein molecule will generate, this figure is on the basis of the quality-charge ratio of protein molecule, shows with the form of the peak figure that is separated from each other.
Because the biomarker in this invention identifies by quality and antibody, thereby they can detect by mass spectroscopy and directly know the identity that they are specific.This method than antibody be the basis ELISA and immunofluorescence technique more accurate.Yet if necessary, these biomarkers also can pass through, such as, determine that amino acid sequence of polypeptide differentiates.For example, a biomarker can be depicted with many enzymes, for example V8 proteinase (V8 protease) or trypsase, and digestion fragment (digestion fragments), claim the polypeptide fingerprint again, molecular weight can be used to search sequence in database, these sequences match with the molecular weight of the digestion segment that is generated by plurality of enzymes.Perhaps, if this biomarker is not the protein molecule in the given data storehouse, on the basis of the N of biomarker utmost point amino acid sequence (N-terminal AminoAcid Sequence), can use the degraded probe, then, these probes can be used to describe by genome that sample generated that has detected biomarker or cDNA storehouse.At last, protein biomarker available protein scalariform ranking method (protein ladder sequencing) sorts.By after molecule being broken into fragment and the method that fragment can be in order removed a single amino acids molecule from the fragment end with enzymolysis or other being handled, can generate protein gradient (protein ladders).Then, with mass spectrum this gradient is analyzed.Stepped fragment (ladder fragments) can identify the amino acid that is removed from molecular end in qualitative difference.Therefore, the present invention can be used for the goldstandard that multiple disease marker is identified.Say that for example if with anti-syphilis antibody (catching specificity syphilis 17kDa albumen), mass spectrogram shows a 17kDa albumen, we can diagnose syphilis; If a 10kDa albumen then may be syphilis 17kDa protein degradation product; If a 30kDa albumen then may be nonspecific proteins.At this moment just can do amino acid sequence with mass spectrum identifies: i.e. the most accurate discriminating (goldstandard).
Embodiment
The differentiation of embodiment 1 normal person and multiple disease antigenic mark in blood
(1) experimental technique
One, material
1. sample is originated: the blood serum sample of collecting the clinical diagnosis patient.50 routine control groups (normal person, known human chorionic gonadtropin, alpha-fetoglobulin, thyrotropic hormone, lactic dehydrogenase, creatine kinase isozyme, the L-aminobutanedioic acid transferase, transferrins, fibrinogen are all normal); The concurrent thyroid gland of 15 routine pregnant woman is hyperfunction, heart failure or infraction (known human chorionic gonadtropin, alpha-fetoglobulin, thyrotropic hormone, lactic dehydrogenase, creatine kinase isozyme, L-aminobutanedioic acid transferase, transferrins, fibrinogen all raises); 16 routine infractions (only lactic dehydrogenase, creatine kinase isozyme raise); The hyperfunction patient of 14 routine thyroid glands (only thyrotropic hormone, transferrins raise); 12 routine normal pregnancies (only human chorionic gonadtropin rising); 15 routine hepatocarcinoma patients (fibrinogen raises for alpha-fetoglobulin only, L-aminobutanedioic acid transferase).
2. reagent: Protein A and G, hcg antibody, alpha-fetoglobulin antibody, thyrotropic hormone antibody, lactate dehydrogenase antibody, creatine kinase isozyme antibody, L-aminobutanedioic acid transferase antibody, transferrins antibody, fibrin original antibody, acetonitrile, trifluoroacetic acid, SINAPINIC acid (Sinapinic acid) are all available from Sigma company or give.
Two, method
1. the collection of sample: draw serum after the whole blood collection, place-80 ℃ of preservations;
2. the preparation of sample: normal human serum; The concurrent thyroid gland of pregnant woman is hyperfunction, heart failure or infraction positive serum; The infraction positive serum; The hyperfunction patient's positive serum of thyroid gland; The normal pregnancies positive serum; The hepatocarcinoma patient positive serum.
3. The pretreatment: earlier 8 kinds of antibody are bonded to simultaneously a Protein A pillar (Column) and go up (the multiple antibody of pillar), then 100 μ l diluted sample are loaded in the pillar and slightly shook at ambient temperature 0.5 hour.From pillar, remove damping fluid (pH7.0-7.4).The same binding buffer liquid of 200 μ l is added in each pillar and concussion was washed 5 minutes.Repeated washing.Removing damping fluid and adding reductive agent (pH3.0-5.0) makes antigen separate from pillar.Get SINAPINIC acid (5mg/mL 50% acetonitrile that supernatant (containing antigen) adds 0.5 μ l; 0.5% trifluoroacetic acid), the drying of giving free rein to.
4. sample detection: sample is put into mass spectrum, will generate flight time mass spectrum.The outside peptide molecule quality standard of using is come the correction mass accuracy.
(2) experimental result
Analyze the composition of protein in 72 routine patients and 50 routine control group (normal person) serum, found that the antigenic mark that 8 protein are formed can divide into groups control group (normal person) with the patient accurately.
Predictablity rate: in statistical analysis and double blinding analysis, the result shows: the antigenic mark of forming with these 8 protein detects, and finds that 72 routine patients and 50 routine control groups (normal person) are correctly divided into groups, and accuracy rate is 100% (122/122).Sensitivity is 100%, and (the concurrent thyroid gland of 15/15 routine pregnant woman is hyperfunction, heart failure or infraction; 16/16 routine infraction; The hyperfunction patient of 14/14 routine thyroid gland; 12/12 routine normal pregnancies; 15/15 routine hepatocarcinoma patient; Correctly divided into groups); Specificity was 100% (50/50 routine control group normal person is correctly divided into groups).
(3) conclusion
To compare from sample with statistical significance patient group and control group (normal specimens), 5 kinds of diseases can be differentiated simultaneously with 8 kinds of serum antigen marks that specific antibody absorption surface material is caught, promptly multiple disease can be differentiated simultaneously with the Mass Spectrometer Method method of these antigenic mark combinations.
In theory, when the specific antibody amount that is added on the antibody absorption surface material ad infinitum increases, the antigenic mark molecule of measuring or biomarker also can correspondingly increase, promptly detect the biomarker more than three kinds simultaneously.On the absorption surface of a multiple antibody, catch multiple antigenic mark, and the antigenic mark of catching is carried out mass spectrophotometry, can detect a plurality of or multiple antigenic mark group simultaneously.
Claims (10)
1. catch the protein analysis method of antigenic mark in the biological sample with antibody absorption surface material for one kind, it is characterized in that adopting mass spectrophotometry that protein group in the sample is differentiated detection.
2. the described protein analysis method of claim 1, the method that wherein said discriminating detects protein group is a mass spectroscopy.
3. the described protein analysis method of claim 1, used antibody absorption surface material is any material that can combine with antibody selectivity or specificity, as albumin A and G (Protein A and Protein G).
4. the described antibody absorption surface of claim 3 material is monoclonal or the polyclonal antibody that is used for the capture antigen mark.
5. the described protein analysis method of claim 1, used antibody absorption surface material is in conjunction with a plurality of or organize monoclonal antibody more and remove to catch a plurality of or multiple biomarker (antigenic mark), can increase the monoclonal antibody group endlessly and reach and detect a plurality of or multiple biomarker or antigenic mark endlessly.
6. the described protein analysis method of claim 1, the analyte that used antibody absorption surface material is caught is biological sample-blood, body fluid, secretion, cytolysate, histolysate and organ dissolved matter.
In the methods analyst biological sample with claim 6 special biomarker to detect the purposes of multiple disease antigen mark.
8. with claim 7, can detect the authentication method of the antigenic mark more than three kinds simultaneously.
9. the described biomarker of claim 5 derives from blood, body fluid, secretion, cytolysis thing, histolysate and organ dissolved matter.
10. the holder of antibody absorption surface material can be sheet metal, glass sheet, potsherd, ceramic bead, magnetic particle or polymer in the claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410004403 CN1657941A (en) | 2004-02-19 | 2004-02-19 | Use of investigating multiple antigen label combined by antibody absorption surface material and mass spectrum |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410004403 CN1657941A (en) | 2004-02-19 | 2004-02-19 | Use of investigating multiple antigen label combined by antibody absorption surface material and mass spectrum |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1657941A true CN1657941A (en) | 2005-08-24 |
Family
ID=35007568
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200410004403 Pending CN1657941A (en) | 2004-02-19 | 2004-02-19 | Use of investigating multiple antigen label combined by antibody absorption surface material and mass spectrum |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1657941A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008043256A1 (en) * | 2006-10-08 | 2008-04-17 | Yang Xu | Test kit for detecting variant or modified biomarker groups with antibody groups and mass spectrometry, and method thereof |
| CN105209907A (en) * | 2013-06-05 | 2015-12-30 | Dh科技发展私人贸易有限公司 | SWATH™ Data Independent Acquisition Technology for Detection of Host Cell Protein Contaminants in Biotherapeutic Protein Products |
-
2004
- 2004-02-19 CN CN 200410004403 patent/CN1657941A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008043256A1 (en) * | 2006-10-08 | 2008-04-17 | Yang Xu | Test kit for detecting variant or modified biomarker groups with antibody groups and mass spectrometry, and method thereof |
| CN101158666B (en) * | 2006-10-08 | 2012-06-27 | 许洋 | Antibody group and mass spectrometric detection variation or modifying biological indication marks group kit and method |
| CN105209907A (en) * | 2013-06-05 | 2015-12-30 | Dh科技发展私人贸易有限公司 | SWATH™ Data Independent Acquisition Technology for Detection of Host Cell Protein Contaminants in Biotherapeutic Protein Products |
| CN105209907B (en) * | 2013-06-05 | 2019-08-16 | Dh科技发展私人贸易有限公司 | SWATH™ Data Independent Acquisition Technology for Detection of Host Cell Protein Contaminants in Biotherapeutic Protein Products |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wolff et al. | Monitoring antibody response following SARS-CoV-2 infection: diagnostic efficiency of 4 automated immunoassays | |
| Mahmood et al. | Raman spectral analysis for rapid screening of dengue infection | |
| CN111443072A (en) | Raman chip for virus detection, preparation method and virus rapid detection method | |
| EP2005172A2 (en) | Apolipoprotein fingerprinting technique | |
| JP2000513436A (en) | Affinity arrays with a wide range of specificities: quantitative methods for the identification of complex samples | |
| RU2397178C1 (en) | Diagnostic test system in immunochip format and differential serum diagnostics of syphilis | |
| CN111423496B (en) | Detection of novel coronavirus polypeptides or combinations thereof | |
| CN105759057A (en) | Dry-type immunofluorescence kit for detecting Alzheimer syndrome MS4A6A and preparation method thereof | |
| US20210046473A1 (en) | Multiplexed mass and nanoparticle detection imaging, tools, fluidics, and methods of making and using the same | |
| Nishino et al. | Article reviewed: Plasma orexin-A is lower in patients with narcolepsy | |
| CN101196526A (en) | Mass spectrometry reagent kit and method for rapid tuberculosis diagnosis | |
| CN1657941A (en) | Use of investigating multiple antigen label combined by antibody absorption surface material and mass spectrum | |
| CN101963618B (en) | Method for identifying heterophilic antibody interference in antibody microarray system and antibody microarray chip using same for detecting target antigen | |
| CN101158666B (en) | Antibody group and mass spectrometric detection variation or modifying biological indication marks group kit and method | |
| Kaysheva et al. | Mass spectrometric detection of the amino acid sequence polymorphism of the hepatitis C virus antigen | |
| CN1614423A (en) | Use for combined detecting multiple virus microbe label by biological chip and mass analyzer | |
| CN118389662A (en) | Biomarker for effectively distinguishing different types of depressive disorder related diseases and application thereof | |
| RU2759149C1 (en) | Method for carrying out an enzyme-linked immunosorbent assay for the detection of antibodies in a human biological sample specific to the sars-cov2 human coronavirus, a test system | |
| CN105807067A (en) | Dry-type immunofluorescence kit for detecting NCAM2 (neural cell adhesion molecules 2) of patient suffering from alzheimer's syndromes and preparation method | |
| CN101907601A (en) | Mass spectrum kit for hypertensive pregnancy biochemical marker and preparation method | |
| WO2021205058A1 (en) | Method for determining coronavirus and kit for the same | |
| JP2009210469A (en) | Analytical method for serum protein | |
| CN1869699A (en) | Method of anaphylactogen screening | |
| CN101153872A (en) | Novel reagent kit for detecting and estimating critical patients and method thereof | |
| CN103454428A (en) | Novel immunomic mass spectrometry kit for detecting individualized insulin and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |