CN117147853A - Application of urine IgG receptor FcRn large subunit p51 and polypeptide fragment thereof in allergic diseases - Google Patents
Application of urine IgG receptor FcRn large subunit p51 and polypeptide fragment thereof in allergic diseases Download PDFInfo
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- CN117147853A CN117147853A CN202210114618.5A CN202210114618A CN117147853A CN 117147853 A CN117147853 A CN 117147853A CN 202210114618 A CN202210114618 A CN 202210114618A CN 117147853 A CN117147853 A CN 117147853A
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- China
- Prior art keywords
- large subunit
- igg receptor
- urine
- receptor fcrn
- use according
- Prior art date
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- 101710177940 IgG receptor FcRn large subunit p51 Proteins 0.000 title claims abstract description 36
- 208000026935 allergic disease Diseases 0.000 title claims abstract description 35
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 29
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 29
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 29
- 239000012634 fragment Substances 0.000 title claims abstract description 23
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Abstract
The invention provides application of a urine IgG receptor FcRn large subunit p51 (IgG receptor FcRn large subunit p) and a polypeptide fragment thereof, in particular application of the urine IgG receptor FcRn large subunit p51 and the polypeptide fragment thereof in preparation of preparations for allergic disease diagnosis, differential diagnosis, disease degree judgment, treatment effect evaluation, monitoring, prognosis evaluation, mechanism research and the like. Allergic diseases are also called allergic diseases. The invention proves that compared with healthy people (normal control group), the urine IgG receptor FcRn large subunit p51 and the polypeptide fragment thereof have increased expression in allergic diseases patients. Can be used for various purposes of detection of allergic diseases patients. The invention plays the advantages of noninvasive acquisition of urine specimen, large-scale repeated sampling and convenient preservation, and utilizes the urine specimen to detect the large subunit p51 of the urine IgG receptor FcRn and the polypeptide fragment thereof.
Description
Technical Field
The invention relates to a new application of a urine IgG receptor FcRn large subunit p51 and a polypeptide fragment thereof, in particular to an application of the urine IgG receptor FcRn large subunit p51 and the polypeptide fragment thereof in preparations such as allergic disease diagnosis, differential diagnosis, disease degree judgment, treatment effect evaluation, monitoring, prognosis evaluation, mechanism research and the like.
Background
Allergic diseases are also called allergic diseases, and according to world health organization statistics, the number of people suffering from allergic diseases is huge worldwide, and about one third of people worldwide suffer from allergic diseases. In recent years, due to the remarkable improvement of industrialization degree, remarkable aggravation of environmental pollution and abrupt change of life style of people, the incidence rate of allergic diseases is increased year by year, and serious allergic diseases even endanger lives, become a global public health focus problem.
At present, clinical auxiliary examination for allergic diseases mainly comprises Skin Prick Test (SPT) and serum sIgE detection, but SPT test patients have poor pain compliance and are easily affected by medicines, and the risk of systemic allergic symptoms is increased, so that the auxiliary examination is not suitable for severe allergic constitution, children and the elderly. The sIgE test is also not a noninvasive test, and often requires repeated blood draws from the patient. For improving the life quality and compliance of allergic disease patients and relieving the pains of repeated blood sampling and pricking tests, the diagnosis of auxiliary allergic disease by painless, convenient, quick and easily repeated urine detection is expected to be realized through urine protein or polypeptide research, and a foundation is laid for further research of urine polypeptide detection kit.
The large subunit p51 (IgG receptor FcRn large subunit p, FCGRT) of the IgG receptor FcRn is encoded by FCGRT, can be combined with beta 2 microglobulin and has the function of an IgG binding molecule, and is involved in cRn immunoglobulin receptor-mediated epithelial cell IgG immunoglobulin endocytosis. In the study, the content of FCGRT in urine of allergic disease patients is up-regulated compared with that of healthy groups, and the content of FCGRT in urine is increased.
Compared with the common clinical blood sample, the urine can be completely and noninvasively collected continuously in a large amount; without steady state regulation, more kinds and larger amplitude of changes can be accumulated, and many pathophysiological changes of the body may be manifested in urine. Protein polypeptides with relatively small molecular weights such as hormones and cytokines can be excreted into urine quickly after entering blood, and the probability of being detected in the urine is much higher than that in the blood; prior to urine collection, the possible proteolytic processing of urine is completed, so that urine proteins remain stable for a longer period of time. In order to relieve the pain of patients with allergic diseases caused by repeated blood sampling, the experiment is expected to realize painless, convenient, quick and easily repeated urine detection to assist diagnosis and disease monitoring of the patients with allergic diseases through urine protein or polypeptide research on the basis of early-stage methodology, and lays a foundation for further research of a urine polypeptide detection kit.
Disclosure of Invention
The invention aims to provide an application of a large subunit p51 of a urine IgG receptor FcRn and a polypeptide fragment thereof in preparation of preparations for allergic disease diagnosis, differential diagnosis, disease degree judgment, treatment effect evaluation, monitoring, prognosis evaluation, mechanism research and the like.
Preferably, the amino acid sequence of the urine IgG receptor FcRn large subunit p51 comprises the amino acid sequence shown in SEQ ID No.1 (MGVPRPQPWA LGLLLFLLPG SLGAESHLSL LYHLTAVSSP APGTPAFWVS GWLGPQQYLS YNSLRGEAEP CGAWVWENQV SWYWEKETTD LRIKEKLFLE AFKALGGKGP YTLQGLLGCE LGPDNTSVPT AKFALNGEEF MNFDLKQGTW GGDWPEALAI SQRWQQQDKA ANKELTFLLF SCPHRLREHL ERGRGNLEWK EPPSMRLKAR PSSPGFSVLT CSAFSFYPPE LQLRFLRNGL AAGTGQGDFG PNSDGSFHAS SSLTVKSGDE HHYCCIVQHA GLAQPLRVEL ESPAKSSVLV VGIVIGVLLL TAAAVGGALL WRRMRSGLPA PWISLRGDDT GVLLPTPGEA QDADLKDVNV IPATA); or an amino acid sequence derived from the amino acid sequence shown in SEQ ID NO.1 and having the same function as the amino acid sequence shown in SEQ ID NO. 1.
Preferably, the preparation is a detection kit for the urine IgG receptor FcRn large subunit p51 and polypeptide fragments thereof of allergic diseases patients.
Preferably, the kit comprises one or more of an immunization method of antigen-antibody reaction and a kit thereof, such as an aptamer antibody or antibody fragment capable of specifically binding to the IgG receptor FcRn large subunit p51 and polypeptide fragments thereof.
Preferably, the detection method comprises a method for directly detecting mass spectrum of the FcRn large subunit p51 of the IgG receptor and polypeptide fragments thereof, and the like and a related kit thereof.
Preferably, the detection method comprises methods such as related nucleic acid detection for directly detecting the FcRn large subunit p51 of the IgG receptor and polypeptide fragments thereof, and related kits thereof.
Preferably, the kit further comprises a component selected from the group consisting of: solid phase carrier, diluent, reference substance, standard substance, quality control substance, detection antibody, secondary antibody diluent, luminous reagent, washing liquid, color developing liquid and stop solution.
Preferably, the standard comprises an IgG receptor FcRn large subunit p51 standard, a humanized tag antibody standard; preferably, the quality control product comprises: igG receptor FcRn large subunit p51 quality control, humanized tag antibody quality control; preferably, the solid support comprises: microparticles, microspheres, slides, test strips, plastic beads, liquid phase chips, microwell plates or affinity membranes, and other carriers of equivalent function.
Preferably, the solid phase carrier is made of any one of polyvinyl chloride, polystyrene, polyacrylamide and cellulose and has similar functions.
The inventor firstly collects urine samples of healthy people and allergic disease patients, after centrifugation for 10min at 2000g, the supernatant is sucked, the concentration of the extracted protein is measured by a Bradford method, and after enzymolysis, the samples are desalted by a C18 desalting column. Adjusting the mass spectrum chromatographic conditions of the DDA library-building sample, adopting skyline software for mass spectrum analysis and library building, adjusting the PRM acquisition mass spectrum parameter setting, completing the integral quantification of the PRM peptide fragment, and processing the data through quantitative value correction/calculation of absolute quantitative values, differential analysis and target protein global cluster analysis. And (3) quantitatively calculating data obtained in mass spectra of the allergic disease group and the normal control group, and finally obtaining the differential protein IgG receptor FcRn large subunit p51.
Compared with healthy people, the invention proves that the IgG receptor FcRn large subunit p51 and the polypeptide fragment thereof are highly expressed in urine of allergic disease patients, and have better consistency with clinical diagnosis. Therefore, the application of the preparation for detecting the large subunit p51 of the urine IgG receptor FcRn and the polypeptide fragment thereof in the diagnosis, differential diagnosis, disease degree judgment, treatment effect evaluation, monitoring, prognosis evaluation, mechanism research and the like of allergic diseases is proposed.
The invention plays the advantages of noninvasive acquisition of urine specimen, large-scale repeated sampling and convenient preservation, and utilizes the urine specimen to detect the large subunit p51 of the urine IgG receptor FcRn and the polypeptide fragment thereof.
The foregoing and other objects, features and advantages of the invention will be apparent from the following more particular description of preferred embodiments, as illustrated in the accompanying drawings.
Drawings
FIG. 1 is a graph showing the content of the large subunit p51 of the urine IgG receptor FcRn and the polypeptide fragment thereof in the allergic disease group and the healthy control group.
Detailed Description
Example 1Urine specimen collection and processing
Allergic disease patients are selected as allergic disease groups, and synchronous healthy physical examination patients are selected as normal control groups. 30ml of fresh morning urine samples of each group of study subjects were collected after admission, and the urine in the morning catheters was collected by the disabled and placed in a dry, clean container. After 2000g of the collected urine specimen is centrifuged for 5min, the supernatant is sucked, each tube is divided into 2ml, 10ul of the supernatant is reserved for protein quantification by adopting a Bradford method, and the supernatant is subjected to enzymolysis and desalting, fraction separation and freeze-drying and then is put on a machine.
Example 2Mass spectrometry and library searching process
Adjusting mass spectrum chromatographic conditions of a DDA library-building sample, adopting skyline software to build a library, selecting unique peptide fragments aiming at each target protein, deriving a unique peptide fragment ion pair list, performing PRM targeted acquisition on the unique peptide fragments, adjusting PRM acquisition mass spectrum parameter setting, and performing integral quantification on the PRM peptide fragments.
Example 3Differential polypeptide data analysis
The data are quantitatively corrected, absolute quantitative values are calculated, FC values and P values of each comparison mode are analyzed and calculated for different comparison groups, and according to screening rules that FC is more than 1.2 or FC is less than 1/1.2 and P is less than 0.05, differential proteins are screened, and global cluster analysis is carried out on target proteins.
Compared with healthy people, the IgG receptor FcRn large subunit p51 is highly expressed in urine of allergic disease patients, the content of the IgG receptor FcRn large subunit p51 in urine of healthy control group and allergic disease group is shown in figure 1, and the expression of the IgG receptor FcRn large subunit p51 in urine of normal control group and allergic disease group has significant difference.
Although the invention has been described with respect to the preferred embodiments, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention, and the invention is defined in the following claims.
Sequence listing
<110> Bowman
<120> use of the large subunit p51 of the urine IgG receptor FcRn and polypeptide fragments thereof in allergic diseases
<130> 22PFCGRT -CN
<140> 22PFCGRT -CN
<141> 2022-01-20
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 365
<212> PRT
<213> Human Urine
<400> 1
Met Gly Val Pro Arg Pro Gln Pro Trp Ala Leu Gly Leu Leu Leu Phe
1 5 10 15
Leu Leu Pro Gly Ser Leu Gly Ala Glu Ser His Leu Ser Leu Leu Tyr
20 25 30
His Leu Thr Ala Val Ser Ser Pro Ala Pro Gly Thr Pro Ala Phe Trp
35 40 45
Val Ser Gly Trp Leu Gly Pro Gln Gln Tyr Leu Ser Tyr Asn Ser Leu
50 55 60
Arg Gly Glu Ala Glu Pro Cys Gly Ala Trp Val Trp Glu Asn Gln Val
65 70 75 80
Ser Trp Tyr Trp Glu Lys Glu Thr Thr Asp Leu Arg Ile Lys Glu Lys
85 90 95
Leu Phe Leu Glu Ala Phe Lys Ala Leu Gly Gly Lys Gly Pro Tyr Thr
100 105 110
Leu Gln Gly Leu Leu Gly Cys Glu Leu Gly Pro Asp Asn Thr Ser Val
115 120 125
Pro Thr Ala Lys Phe Ala Leu Asn Gly Glu Glu Phe Met Asn Phe Asp
130 135 140
Leu Lys Gln Gly Thr Trp Gly Gly Asp Trp Pro Glu Ala Leu Ala Ile
145 150 155 160
Ser Gln Arg Trp Gln Gln Gln Asp Lys Ala Ala Asn Lys Glu Leu Thr
165 170 175
Phe Leu Leu Phe Ser Cys Pro His Arg Leu Arg Glu His Leu Glu Arg
180 185 190
Gly Arg Gly Asn Leu Glu Trp Lys Glu Pro Pro Ser Met Arg Leu Lys
195 200 205
Ala Arg Pro Ser Ser Pro Gly Phe Ser Val Leu Thr Cys Ser Ala Phe
210 215 220
Ser Phe Tyr Pro Pro Glu Leu Gln Leu Arg Phe Leu Arg Asn Gly Leu
225 230 235 240
Ala Ala Gly Thr Gly Gln Gly Asp Phe Gly Pro Asn Ser Asp Gly Ser
245 250 255
Phe His Ala Ser Ser Ser Leu Thr Val Lys Ser Gly Asp Glu His His
260 265 270
Tyr Cys Cys Ile Val Gln His Ala Gly Leu Ala Gln Pro Leu Arg Val
275 280 285
Glu Leu Glu Ser Pro Ala Lys Ser Ser Val Leu Val Val Gly Ile Val
290 295 300
Ile Gly Val Leu Leu Leu Thr Ala Ala Ala Val Gly Gly Ala Leu Leu
305 310 315 320
Trp Arg Arg Met Arg Ser Gly Leu Pro Ala Pro Trp Ile Ser Leu Arg
325 330 335
Gly Asp Asp Thr Gly Val Leu Leu Pro Thr Pro Gly Glu Ala Gln Asp
340 345 350
Ala Asp Leu Lys Asp Val Asn Val Ile Pro Ala Thr Ala
355 360 365
Claims (9)
1. The application of the large subunit p51 of the urine IgG receptor FcRn and the polypeptide fragment thereof in preparing preparations for allergic disease diagnosis, differential diagnosis, disease degree judgment, treatment effect evaluation, monitoring, prognosis evaluation, mechanism research and the like.
2. The use according to claim 1, wherein the amino acid sequence of the large subunit p51 of FcRn of the urinary IgG receptor is shown in SEQ ID No. 1; or an amino acid sequence derived from the amino acid sequence shown in SEQ ID NO.1 and having the same function as the amino acid sequence shown in SEQ ID NO. 1.
3. The use according to claim 1, wherein the preparation is a kit for detecting the large subunit p51 of FcRn, the IgG receptor in urine of patients suffering from allergic diseases, and polypeptide fragments thereof.
4. The use according to claim 3, wherein the kit comprises one or more of an immunization method of an antigen-antibody reaction and a kit thereof such as an aptamer antibody or antibody fragment capable of specifically binding to FcRn large subunit p51 of IgG receptor and polypeptide fragments thereof.
5. The use according to claim 3, wherein the detection method comprises a method for directly detecting mass spectra of the large subunit p51 of the IgG receptor FcRn and polypeptide fragments thereof, and the like, and a kit related thereto.
6. The use according to claim 3, wherein the detection method comprises a method for directly detecting the large subunit p51 of the IgG receptor FcRn and polypeptide fragments thereof or related nucleic acid detection and the like and related kit thereof.
7. The use according to claim 3, wherein the kit further comprises a component selected from the group consisting of: solid phase carrier, diluent, reference substance, standard substance, quality control substance, detection antibody, secondary antibody diluent, luminous reagent, washing liquid, color developing liquid and stop solution.
8. The use according to claim 7, wherein the standard comprises an IgG receptor FcRn large subunit p51 standard, a humanized tag antibody standard; preferably, the quality control product comprises: igG receptor FcRn large subunit p51 controls, humanized tag antibody controls; preferably, the solid support comprises: microparticles, microspheres, slides, test strips, plastic beads, liquid phase chips, microwell plates or affinity membranes, and other carriers of equivalent function.
9. The use according to claim 8, wherein the solid support is made of any one of polyvinyl chloride, polystyrene, polyacrylamide, cellulose and a support with similar function.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210114618.5A CN117147853A (en) | 2022-01-30 | 2022-01-30 | Application of urine IgG receptor FcRn large subunit p51 and polypeptide fragment thereof in allergic diseases |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210114618.5A CN117147853A (en) | 2022-01-30 | 2022-01-30 | Application of urine IgG receptor FcRn large subunit p51 and polypeptide fragment thereof in allergic diseases |
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