CN113267587B - Characteristic peptide fragment and method for measuring content of pro-SFTPB standard substance - Google Patents
Characteristic peptide fragment and method for measuring content of pro-SFTPB standard substance Download PDFInfo
- Publication number
- CN113267587B CN113267587B CN202110606380.3A CN202110606380A CN113267587B CN 113267587 B CN113267587 B CN 113267587B CN 202110606380 A CN202110606380 A CN 202110606380A CN 113267587 B CN113267587 B CN 113267587B
- Authority
- CN
- China
- Prior art keywords
- pro
- sftpb
- peptide
- protein
- standard
- 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.)
- Active
Links
- 108010033276 Peptide Fragments Proteins 0.000 title claims abstract description 69
- 102000007079 Peptide Fragments Human genes 0.000 title claims abstract description 67
- 101001086862 Homo sapiens Pulmonary surfactant-associated protein B Proteins 0.000 title claims abstract description 49
- 102100032617 Pulmonary surfactant-associated protein B Human genes 0.000 title claims abstract description 49
- 239000000126 substance Substances 0.000 title claims abstract description 29
- 238000000034 method Methods 0.000 title claims abstract description 24
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 39
- 102000004169 proteins and genes Human genes 0.000 claims description 34
- 108090000623 proteins and genes Proteins 0.000 claims description 34
- 239000000243 solution Substances 0.000 claims description 18
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 claims description 9
- 238000004458 analytical method Methods 0.000 claims description 7
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 claims description 6
- 238000006243 chemical reaction Methods 0.000 claims description 6
- 239000012895 dilution Substances 0.000 claims description 5
- 238000010790 dilution Methods 0.000 claims description 5
- 239000004475 Arginine Substances 0.000 claims description 4
- 239000004472 Lysine Substances 0.000 claims description 4
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims description 4
- PZBFGYYEXUXCOF-UHFFFAOYSA-N TCEP Chemical compound OC(=O)CCP(CCC(O)=O)CCC(O)=O PZBFGYYEXUXCOF-UHFFFAOYSA-N 0.000 claims description 4
- 230000029936 alkylation Effects 0.000 claims description 4
- 238000005804 alkylation reaction Methods 0.000 claims description 4
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 4
- 238000004108 freeze drying Methods 0.000 claims description 4
- ATRRKUHOCOJYRX-UHFFFAOYSA-N Ammonium bicarbonate Chemical compound [NH4+].OC([O-])=O ATRRKUHOCOJYRX-UHFFFAOYSA-N 0.000 claims description 3
- 229910000013 Ammonium bicarbonate Inorganic materials 0.000 claims description 3
- 102000004142 Trypsin Human genes 0.000 claims description 3
- 108090000631 Trypsin Proteins 0.000 claims description 3
- 235000012538 ammonium bicarbonate Nutrition 0.000 claims description 3
- 239000001099 ammonium carbonate Substances 0.000 claims description 3
- 238000004364 calculation method Methods 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- PGLTVOMIXTUURA-UHFFFAOYSA-N iodoacetamide Chemical compound NC(=O)CI PGLTVOMIXTUURA-UHFFFAOYSA-N 0.000 claims description 3
- 239000012460 protein solution Substances 0.000 claims description 3
- 238000006722 reduction reaction Methods 0.000 claims description 3
- 239000012588 trypsin Substances 0.000 claims description 3
- 238000002372 labelling Methods 0.000 claims 2
- 238000004750 isotope dilution mass spectroscopy Methods 0.000 abstract description 3
- 230000009286 beneficial effect Effects 0.000 abstract description 2
- 238000003908 quality control method Methods 0.000 abstract description 2
- 238000013441 quality evaluation Methods 0.000 abstract description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 10
- 239000013582 standard series solution Substances 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- 239000012086 standard solution Substances 0.000 description 6
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 5
- PBCJIPOGFJYBJE-UHFFFAOYSA-N acetonitrile;hydrate Chemical compound O.CC#N PBCJIPOGFJYBJE-UHFFFAOYSA-N 0.000 description 5
- 235000019253 formic acid Nutrition 0.000 description 5
- 238000002156 mixing Methods 0.000 description 5
- 239000000843 powder Substances 0.000 description 4
- 239000012224 working solution Substances 0.000 description 4
- 108010041520 Pulmonary Surfactant-Associated Proteins Proteins 0.000 description 3
- 102000000528 Pulmonary Surfactant-Associated Proteins Human genes 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 238000012544 monitoring process Methods 0.000 description 3
- 238000011002 quantification Methods 0.000 description 3
- 238000007792 addition Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 229940066294 lung surfactant Drugs 0.000 description 2
- 239000003580 lung surfactant Substances 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 108010042653 IgA receptor Proteins 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 102100034014 Prolyl 3-hydroxylase 3 Human genes 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The application discloses a characteristic peptide segment and a method for measuring the content of pro-SFTPB standard substances, wherein the characteristic peptide segment for measuring the content of the pro-SFTPB standard substances is at least one selected from DPLPDPLLDK and LVLPVLPGALQAR; the beneficial effects of the application are as follows: the application provides a plurality of pro-SFTPB characteristic peptide fragments, and applies the characteristic peptide fragments to the determination of the content of pro-SFTPB standard substances for the first time, so that a high-sensitivity and high-selectivity pro-SFTPB isotope dilution mass spectrometry method is established, the result can be traced to SI units finally, the blank of the prior art is filled, and the method has important significance for establishing a quality control and evaluation system of the pro-SFTPB standard substances.
Description
Technical Field
The application relates to a protein analysis and detection technology, in particular to a characteristic peptide segment and a method for determining the content of a lung surfactant protein B precursor (pro-SFTPB) standard substance.
Background
The lung surfactant protein B precursor (Pro-surfactant protein B, pro-SFTPB) can promote the modification and maturation of the lung surfactant protein B, regulate and maintain the internal environment stability of lung surfactant substances, promote the reuse of lung surfactant lipid and protein, and can be applied to early screening diagnosis and prognosis monitoring of cancers.
Whether the quantitative determination result of pro-SFTPB in clinical detection is reliable depends on the accuracy of the quantitative determination of the protein standard substance, but no determination method of the content of the pro-SFTPB standard substance exists at present. In view of the above-mentioned needs, the present application provides a characteristic peptide fragment for determining the content of pro-SFTPB standard substance, and a specific determination method. The method can trace to SI units finally, and has good accuracy and reliability.
Disclosure of Invention
The main purpose of the application is to provide a characteristic peptide fragment and a method for measuring the content of pro-SFTPB standard substances.
In order to achieve the above object, the present application provides the following technical solutions:
the characteristic peptide fragment for measuring the content of pro-SFTPB standard substance is at least one selected from DPLPDPLLDK (hereinafter referred to as SFTPBa) and LVLPVLPGALQAR (hereinafter referred to as SFTPBb).
The above-mentioned characteristic peptide fragment for measuring the content of pro-SFTPB standard substance is, as a preferred embodiment, a peptide fragment of the characteristic peptide fragment SFTPBa wherein the C and N elements on the lysine are respectively 13 C and C 15 N is marked to obtain corresponding internal standard peptide DPLPDPLLDK # -, and 13 C 6 , 15 N 2 ) (hereinafter referred to as SFTPBa_IS).
The above-mentioned characteristic peptide fragment for measuring the content of pro-SFTPB standard substance is, as a preferred embodiment, a fragment of characteristic peptide SFTPBb wherein the C and N elements on arginine are respectively 13 C and C 15 N is marked to obtain corresponding internal standard peptide LVLPVLPGALQAR # -, and 13 C 6 , 15 N 4 ) (hereinafter referred to as SFTPBb_IS).
In a second aspect of the present application, there is provided a method for determining the content of pro-SFTPB standard substances, wherein a pro-SFTPB standard protein sample is subjected to reduction, alkylation, enzymolysis, drying, adding the internal standard peptide fragment, and then performing nano liter liquid chromatography-mass spectrometry analysis to quantify the pro-SFTPB protein.
The method described above, as a preferred embodiment, the establishment of the standard curve, comprises the steps of: mixing characteristic peptide fragment solutions of pro-SFTPB with different concentrations with an internal standard peptide fragment solution with fixed concentration in equal volume to prepare a standard series solution, then carrying out nano liter liquid chromatography-mass spectrometry analysis, and establishing a standard curve by taking the concentration of the characteristic peptide fragment as an abscissa and the peak area ratio of the characteristic peptide fragment and the internal standard peptide fragment as an ordinate;
the method comprises the following specific steps:
taking freeze-dried powder of the characteristic peptide fragment of pro-SFTPB and the internal standard peptide fragment, and dissolving the freeze-dried powder with 5% acetonitrile water solution; SFTPBa and SFTPBb were prepared as mixed standard solutions of equal concentration, and diluted with 15% acetonitrile aqueous solution (containing 0.1% formic acid) to 0.5ng/mL, 2.0ng/mL, 5.0ng/mL, 20.0ng/mL and 50.0ng/mL, respectively, to obtain mixed standard series solutions.
Simultaneously preparing SFTPBa_IS and SFTPBb_IS into mixed internal standard solution with equal concentration, and diluting the mixed internal standard solution into mixed internal standard working solution with 15 percent of acetonitrile water solution (containing 0.1 percent of formic acid) to 5.0 ng/mL;
mixing the mixed standard series solutions of pro-SFTPB with different concentrations with 5.0ng/mL mixed internal standard working solution in equal volume to prepare a standard series solution, then carrying out nano liter liquid chromatography-mass spectrometry, collecting data in a parallel reaction monitoring (Parallel Reaction Monitoring, PRM) scanning mode, and quantitatively analyzing by Skyline software; and drawing a standard curve by taking the concentration of the characteristic peptide fragment as an abscissa and the peak area ratio of the characteristic peptide fragment and the internal standard peptide fragment as an ordinate and the weight as 1/x.
In the method, as a preferred implementation scheme, a certain volume of pro-SFTPB standard protein solution is accurately measured, TCEP is added for reduction reaction, iodoacetamide is then added for alkylation, and then ammonium bicarbonate solution of trypsin is added for enzymolysis; after enzymolysis, adding trifluoroacetic acid to terminate the enzymolysis reaction; drying a sample, adding an internal standard peptide fragment solution, and carrying out nano liter liquid chromatography-mass spectrometry analysis; peak area of characteristic peptide fragment and internal standard peptide fragment in sampleSubstituting the ratio into a standard curve to obtain the concentration C of the characteristic peptide fragment peptide， And calculating the concentration of the pro-SFTPB protein standard substance.
protein concentration C of pro-SFTPB standard substance protein The calculation formula of (2) is as follows:
C protein ＝C peptide ×V Dilution ×R mol ×M protein /V sample ×M peptide ；
wherein:
C protein representing the protein concentration in the original protein sample;
C peptide representing the concentration of the characteristic peptide fragments measured in the sample of the machine;
V Dilution representing the volume of the sample after freeze-drying and reconstitution;
R mol representing the molar ratio of the measured protein to the measured characteristic peptide fragment;
M protein indicating the molecular weight of the protein measured;
V sample representing the volume of the original protein sample;
M peptide representing the molecular weight of the characteristic peptide fragment measured.
Compared with the prior art, the application has the beneficial effects that:
(1) The application provides a plurality of pro-SFTPB characteristic peptide fragments, and applies the characteristic peptide fragments to the determination of the content of pro-SFTPB standard substances for the first time, so that a high-sensitivity and high-selectivity pro-SFTPB isotope dilution mass spectrometry method is established, the result can be traced to SI units finally, the blank of the prior art is filled, and the method has important significance for establishing a quality control and evaluation system of the pro-SFTPB standard substances.
(2) When the isotope dilution mass spectrometry is adopted to quantify the pro-SFTPB standard substance, the average value of the quantification results of two characteristic peptide fragments can be selected at the same time, and the accuracy and the reliability of the quantification results are improved.
Drawings
FIG. 1 is a standard curve of a characteristic peptide segment SFTPBa;
FIG. 2 is a standard curve of a characteristic peptide segment SFTPBb;
FIG. 3 is a chromatogram of a characteristic peptide fragment SFTPBa;
FIG. 4 IS a chromatogram of an internal standard peptide segment SFTPBa_IS;
FIG. 5 is a chromatogram of a characteristic peptide fragment SFTPBb;
FIG. 6 IS a chromatogram of the internal standard peptide fragment SFTPBb_IS.
Detailed Description
In order that those skilled in the art will better understand the present application, a technical solution of embodiments of the present application will be clearly and completely described in the following description with reference to examples, and it is apparent that the described embodiments are only some embodiments of the present application, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the present application without making any inventive effort, shall fall within the scope of the present application.
The application provides a characteristic peptide segment for measuring the content of a pro-SFTPB standard substance, which is selected from at least one of DPLPDPLLDK and LVLPVLPGALQAR;
the C and N elements on the lysine of the characteristic peptide DPLPDPLLDK are respectively carried out 13 C and C 15 N is marked to obtain the corresponding internal standard peptide DPLPDPLLDK 13 C 6 , 15 N 2 The method comprises the steps of carrying out a first treatment on the surface of the The C and N elements on the arginine of the characteristic peptide LVLPVLPGALQAR are respectively carried out 13 C and C 15 N is marked to obtain the corresponding internal standard peptide LVLPVLPGALQAR 13 C 6 , 15 N 4 。
The method for measuring the content of pro-SFTPB standard substances by adopting the characteristic peptide fragment and the internal standard peptide fragment comprises the following steps:
(1) Selecting two characteristic peptide fragments of pro-SFTPB: DPLPDPLLDK and LVLPVLPGALQAR; DPLPDPLLDK is named SFTPBa, LVLPVLPGALQAR as SFTPBb;
(2) C and N elements on the characteristic peptide SFTPBa lysine are respectively carried out 13 C and C 15 N is marked to obtain corresponding internal standard peptide DPLPDPLLDK # -, and 13 C 6 , 15 N 2 ) Named SFTPBa_IS; for characteristics ofThe C and N elements on the peptide SFTPBb arginine are respectively carried out 13 C and C 15 N is marked to obtain corresponding internal standard peptide LVLPVLPGALQAR # -, and 13 C 6 , 15 N 4 ) Named SFTPBb_IS;
(3) Mixing characteristic peptide fragments with different concentrations and internal standard peptide fragments with fixed concentrations in equal volumes, performing nano liter liquid chromatography-mass spectrometry analysis, and establishing a standard curve;
the standard curve is established by the following steps:
taking freeze-dried powder of the characteristic peptide fragment of pro-SFTPB and the internal standard peptide fragment, and dissolving the freeze-dried powder with 5% acetonitrile water solution; SFTPBa and SFTPBb were prepared as mixed standard solutions of equal concentration, and diluted with 15% acetonitrile aqueous solution (containing 0.1% formic acid) to 0.5ng/mL, 2.0ng/mL, 5.0ng/mL, 20.0ng/mL and 50.0ng/mL, respectively, to obtain mixed standard series solutions.
Simultaneously preparing SFTPBa_IS and SFTPBb_IS into mixed internal standard solution with equal concentration, and diluting the mixed internal standard solution into mixed internal standard working solution with 15 percent of acetonitrile water solution (containing 0.1 percent of formic acid) to 5.0 ng/mL;
mixing the mixed standard series solutions of pro-SFTPB with different concentrations with 5.0ng/mL mixed internal standard working solution in equal volume to prepare a standard series solution, then carrying out nano liter liquid chromatography-mass spectrometry, collecting data (shown in table 1) in a PRM scanning mode, and quantitatively analyzing by Skyline software; drawing a standard curve by taking the concentration of the characteristic peptide fragment as an abscissa and the peak area ratio of the characteristic peptide fragment and the internal standard peptide fragment as an ordinate, and the weight as 1/x; the standard curves of the characteristic peptide fragments are shown in fig. 1 and 2;
in the quantification process, in order to avoid scanning errors of single sub-ions, the extraction chromatographic peak areas of 5 sub-ions of the target peptide fragment are added to be used as the peak area of the target peptide fragment;
TABLE 1 PRM Scan information for Pro-SFTPB
(4) Accurately measuring a certain volume of pro-SFTPB standard protein solution (recording volume V sample ) Adding 0.5mol/L TCEP to make the final concentration of TCEP be 25mmol/L, reacting for 30min at 37 ℃, then adding 2uL iodoacetamide with the concentration of 20mmol/L for alkylation, and reacting for 30min at room temperature in a dark place; a further 40uL of trypsin in ammonium bicarbonate (50 ng/uL in 50mM NH 4 HCO 3 ) Enzymolysis is carried out for 24 hours at 37 ℃; after enzymolysis, adding 4uL of 10% trifluoroacetic acid aqueous solution to terminate the enzymolysis reaction; freeze-drying the sample, re-dissolving with 15% acetonitrile water solution (containing 0.1% formic acid), mixing with the internal standard peptide fragment solution in equal volume, taking 2ul for nano liter liquid chromatography-mass spectrometry analysis, accurately quantifying two characteristic peptide fragments, and recording chromatograms, wherein the chromatograms are shown in figures 3-6;
quantitative analysis is carried out on the collected mass spectrum data through Skyline software, the peak area ratio of the characteristic peptide fragment and the internal standard peptide fragment in the upper sample is substituted into a standard curve, and the concentration C of the characteristic peptide fragment in the upper sample is obtained peptide 。
(5) Calculating the concentration of the pro-SFTPB protein standard substance according to the peptide fragment content of the solution in the step (4);
concentration C of pro-SFTPB protein standard substance protein The calculation formula of (2) is as follows:
C protein ＝C peptide ×V Dilution ×R mol ×M protein /V sample ×M peptide ；
wherein:
C protein representing the protein concentration in the original protein sample;
C peptide representing the concentration of the characteristic peptide fragments measured in the sample of the machine;
V Dilution representing the volume of the sample after freeze-drying and reconstitution;
R mol representing the molar ratio of the measured protein to the measured characteristic peptide fragment;
M protein indicating the molecular weight of the protein measured;
V sample representing the volume of the original protein sample;
M peptide representing the molecular weight of the characteristic peptide fragment measured.
And when the protein concentration is calculated, selecting a plurality of quantitative results of the enzyme digestion peptide fragments for conversion. According to the embodiment of the application, the average value of the quantitative results of two enzyme-cleaved peptide fragments is selected at the same time, and the quantitative result of any peptide fragment can be selected.
The foregoing is merely a preferred embodiment of the present application, and it should be noted that modifications and additions may be made to those skilled in the art without departing from the method of the present application, which modifications and additions are also to be considered as within the scope of the present application.
Claims (4)
1. A method for measuring the content of pro-SFTPB standard substance is characterized in that a certain volume of pro-SFTPB standard protein solution is accurately measured, TCEP is added for reduction reaction, iodoacetamide is then added for alkylation, and then ammonium bicarbonate solution of trypsin is added for enzymolysis; after enzymolysis, adding trifluoroacetic acid to terminate the enzymolysis reaction; drying a sample, adding an internal standard peptide fragment solution, and then carrying out nano liter liquid chromatography-mass spectrometry analysis to quantify a pro-SFTPB standard protein sample;
wherein, the characteristic peptide fragments for measuring the content of the pro-SFTPB standard substance are DPLPDPLLDK and LVLPVLPGALQAR; the internal standard peptide is prepared by respectively carrying out C and N elements on the lysine of the characteristic peptide DPLPDPLLDK 13 C and C 15 N-labelling to obtain the corresponding internal standard peptide DPLPDPLLDK 13 C6, 15 N2, and C and N elements on the arginine of the characteristic peptide LVLPVLPGALQAR, respectively 13 C and C 15 N-labelling to obtain the corresponding internal standard peptide LVLPVLPGALQAR 13 C6, 15 N4; the quantitative result is the average value of the quantitative results of two enzyme-cleaved peptide fragments.
2. The method for determining the content of pro-SFTPB standard substances according to claim 1, wherein characteristic peptide solutions of different concentrations of pro-SFTPB are mixed with internal standard peptide solutions of fixed concentrations in equal volumes to prepare a standard series of solutions, then nano liter liquid chromatography-mass spectrometry is carried out, and a standard curve is established by taking the concentration of the characteristic peptide as an abscissa and the ratio of the peak areas of the characteristic peptide and the internal standard peptide as an ordinate.
3. The method for determining the content of pro-SFTPB standard substance according to claim 2, wherein the peak area ratio of the characteristic peptide fragment and the internal standard peptide fragment in the sample is substituted into the standard curve to determine the concentration C of the characteristic peptide fragment peptide 。
4. The method for determining the content of pro-SFTPB protein according to claim 3, wherein the concentration C of the pro-SFTPB protein is protein The calculation formula of (2) is as follows:
wherein:
C protein representing the pro-SFTPB protein concentration in the original protein sample;
C peptide representing the concentration of the characteristic peptide fragments measured in the sample of the machine;
V Dilution representing the volume of the sample after freeze-drying and reconstitution;
R mol representing the molar ratio of the measured protein to the measured characteristic peptide fragment;
M protein indicating the molecular weight of the protein measured;
V sample representing the volume of the original protein sample;
M peptide representing the molecular weight of the characteristic peptide fragment measured.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110606380.3A CN113267587B (en) | 2021-05-27 | 2021-05-27 | Characteristic peptide fragment and method for measuring content of pro-SFTPB standard substance |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110606380.3A CN113267587B (en) | 2021-05-27 | 2021-05-27 | Characteristic peptide fragment and method for measuring content of pro-SFTPB standard substance |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113267587A CN113267587A (en) | 2021-08-17 |
| CN113267587B true CN113267587B (en) | 2023-11-10 |
Family
ID=77234024
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110606380.3A Active CN113267587B (en) | 2021-05-27 | 2021-05-27 | Characteristic peptide fragment and method for measuring content of pro-SFTPB standard substance |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113267587B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114166966B (en) * | 2021-11-26 | 2024-03-19 | 杭州广科安德生物科技有限公司 | Method for determining pro-SFTPB value in serum |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1997041155A2 (en) * | 1996-04-26 | 1997-11-06 | Ony, Inc. | Compositions and methods for isolating lung surfactant hydrophobic proteins sp-b and sp-c |
| CA2907120A1 (en) * | 2013-03-15 | 2014-09-18 | Sera Prognostics, Inc. | Biomarkers and methods for predicting preterm birth |
| JP2017129459A (en) * | 2016-01-20 | 2017-07-27 | 日本電子株式会社 | Mass spectrometer and mass spectrometry |
| CN110174456A (en) * | 2019-06-11 | 2019-08-27 | 华润双鹤药业股份有限公司 | A kind of method and its application measuring Pulmonary surfactant protein |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IL172297A (en) * | 2005-10-03 | 2016-03-31 | Compugen Ltd | Soluble vegfr-1 variants for diagnosis of preeclamsia |
| EP2021492A2 (en) * | 2006-04-28 | 2009-02-11 | Ludwig Institute For Cancer Research | Quantification of enzyme activity by mass spectrometry |
| US20160060329A1 (en) * | 2014-07-28 | 2016-03-03 | Donald SIN | Compositions and methods for the diagnosis and prognosis of lung cancer |
| CN110291397A (en) * | 2017-02-09 | 2019-09-27 | 得克萨斯大学体系董事会 | The detection and treatment method of lung cancer |
-
2021
- 2021-05-27 CN CN202110606380.3A patent/CN113267587B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1997041155A2 (en) * | 1996-04-26 | 1997-11-06 | Ony, Inc. | Compositions and methods for isolating lung surfactant hydrophobic proteins sp-b and sp-c |
| CA2907120A1 (en) * | 2013-03-15 | 2014-09-18 | Sera Prognostics, Inc. | Biomarkers and methods for predicting preterm birth |
| CN106574919A (en) * | 2013-03-15 | 2017-04-19 | 赛拉预测公司 | Biomarkers and methods for predicting preterm birth |
| JP2017129459A (en) * | 2016-01-20 | 2017-07-27 | 日本電子株式会社 | Mass spectrometer and mass spectrometry |
| CN110174456A (en) * | 2019-06-11 | 2019-08-27 | 华润双鹤药业股份有限公司 | A kind of method and its application measuring Pulmonary surfactant protein |
Non-Patent Citations (5)
| Title |
|---|
| Aberrant processing forms of lung surfactant proteins SP–B and SP–C revealed by high-resolution mass spectrometry;Dmitry Galetskiy et al.;《Eur. J. Mass Spectrom》;20081112;图1 * |
| Daniel J Wegner et al..A major deletion in the surfactant protein-B gene causing lethal respiratory distress.《Acta Pædiatrica》.2007, * |
| Dmitry Galetskiy et al..Aberrant processing forms of lung surfactant proteins SP–B and SP–C revealed by high-resolution mass spectrometry.《Eur. J. Mass Spectrom》.2008, * |
| Measurement of Human Surfactant Protein-B Turnover in Vivo from Tracheal Aspirates Using Targeted Proteomics;Daniela M. Tomazela et al.;《Anal. Chem.》;20101231;用于SP-B示踪物测量的气管吸出物的制备、液相色谱-串联质谱法、肽丰度的量化、选择用于示踪物富集测量的肽转变、讨论、表2 * |
| 罗云波 等.《食品生物技术导论》.中国农业大学出版社,2016,第386页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113267587A (en) | 2021-08-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US6258605B1 (en) | Clinical method for the genetic screening of newborns using tandem mass spectrometry | |
| CN113267587B (en) | Characteristic peptide fragment and method for measuring content of pro-SFTPB standard substance | |
| CN111289425B (en) | Protein content measurement reference method based on fluorescence labeling flow type single molecule counting | |
| US7297545B2 (en) | Clinical method for the genetic screening of newborns using tandem mass spectrometry and internal standards therefor | |
| CN114166966A (en) | Method for determining value of pro-SFTPB in serum | |
| CN114460188A (en) | Kit for detecting human plasma catecholamine intermediate metabolite and testing method | |
| CN115902048A (en) | Method for detecting water-soluble vitamins in serum by methyl derivatization-high performance liquid chromatography tandem mass spectrometry | |
| CN113295805A (en) | Method for detecting hydrazine hydrate in medicine | |
| CN107064265A (en) | A kind of electrochemica biological sensor for being used for HbA1c detections of MPBA modifications and its preparation and application | |
| CN113219117A (en) | Mass spectrometry method of TIMP1 protein standard substance | |
| CN107573536B (en) | Preparation method and application of aerogel | |
| CN108732222B (en) | Method for simultaneously and rapidly detecting glycosylated hemoglobin and glycosylated serum protein in blood | |
| CN1896271A (en) | Reagent determination by serum potassium ion enzyme method | |
| CN108387575A (en) | A method of measuring Plant Total Nitrogen using Continuous Flow Analysis instrument | |
| CN1869652A (en) | Investigating agent and method of serum sodium ion enzyme method | |
| CN110903444B (en) | Polymer silver-coated micro-nano particle and method for detecting urine micromolecules by using same | |
| CN114609295A (en) | High performance liquid chromatography analysis method for quinic acid content in tala enzymolysis waste liquid | |
| CN107764877B (en) | A kind of highly sensitive serum tolal bile acid electrochemical detection method | |
| CN103529199B (en) | The method of clenbuterol content in the animal derived sample of a kind of field quick detection | |
| CN1010614B (en) | Analyzer for local molecular structure | |
| CN200982972Y (en) | Malignancy electrochemical screening and early diagnostic apparatus | |
| CN113281442A (en) | Method for valuing standard substance rich in leucine alpha-2 glycoprotein 1 | |
| CN1866018A (en) | Electrochemical screening and early diagnosing instrument for malignant tumor | |
| CN113466384B (en) | Liquid chromatography tandem mass spectrometry quantitative detection method for content of glycosylated hemoglobin in whole blood | |
| CN113984863B (en) | Glycosylated hemoglobin detection method based on aptamer biosensor |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |