CN117147850A - Application of nerve guidance molecule 5A and polypeptide fragment thereof as urine reference marker - Google Patents

Application of nerve guidance molecule 5A and polypeptide fragment thereof as urine reference marker Download PDF

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CN117147850A
CN117147850A CN202111561840.1A CN202111561840A CN117147850A CN 117147850 A CN117147850 A CN 117147850A CN 202111561840 A CN202111561840 A CN 202111561840A CN 117147850 A CN117147850 A CN 117147850A
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urine
exosome
ser
leu
gly
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张曼
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    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
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    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
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    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54306—Solid-phase reaction mechanisms
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    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
    • G01N33/6848—Methods of protein analysis involving mass spectrometry
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • G01N2800/00—Detection or diagnosis of diseases
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Abstract

The invention provides an application of a urine exosome nerve guiding molecule 5A (Semaphorin-5A, sema 5A) and a polypeptide fragment thereof, in particular to an application of Sema 5A and a polypeptide fragment thereof as urine exosome stable expression protein, which has the characteristic of urine stable expression in the whole life cycle of children, teenagers, middle-aged and elderly people, can be used as an urine internal reference marker and a comparison marker to be applied to urine exosome detection, exosome research, health population clinical research, related disease diagnosis, differential diagnosis, illness state and activity judgment, treatment effect evaluation, monitoring, prognosis evaluation and mechanism research. According to the invention, the research proves that the urine exosome Sema 5A and the polypeptide fragment thereof are stably expressed in urine of the whole human life cycle of children, teenagers, middle-aged and elderly people, and can be used as an internal urine reference marker and a comparison marker for detecting urine exosome, and the exosome research, the clinical research of healthy people, the related disease diagnosis, the differential diagnosis, the disease degree and activity judgment, the treatment effect evaluation, the monitoring, the prognosis evaluation and the various purposes of mechanism research are applied and detected. 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 urine exosome nerve guiding molecule 5A and the polypeptide fragment thereof.

Description

Application of nerve guidance molecule 5A and polypeptide fragment thereof as urine reference marker
Technical Field
The invention relates to a new application of a urine exosome nerve guiding molecule 5A and a polypeptide fragment thereof, in particular to application of the nerve guiding molecule 5A and the polypeptide fragment thereof as a stable expression protein of urine exosome, which has the characteristics of stable expression of urine in the whole life cycle of children, teenagers, middle-aged and elderly people, can be used as an urine internal reference marker and a comparison marker in preparation of urine exosome detection, exosome research, clinical research of healthy people, related disease diagnosis, differential diagnosis, disease degree and activity judgment, treatment effect evaluation, monitoring, prognosis evaluation and mechanism research.
Background
The nerve guidance molecule 5A (Semaphorin-5A, sema 5A) is one of the members of the family of nerve axon guidance molecules, which is originally recognized to a certain extent during the growth and development of the brain nerve, and has the main functions of playing a role in chemical rejection, regulating the growth direction of axons, guiding the axons into the correct positions and establishing synaptic connection. Studies have shown that Sema 5A is involved in a number of pathophysiological processes in humans, such as cell migration, tumor growth, angiogenesis, and immunomodulation. Sema 5A also promotes proliferation of T cells, natural killer cells, and induces secretion of T helper 1/17 inflammatory cytokines. Recent studies have also found high expression of Sema 5A in gastric cancer, glioma, pancreatic cancer, lung cancer, prostate cancer, and melanoma, which may be involved in biological processes that regulate tumorigenesis and formation, and have demonstrated that Sema 5A is closely related to tumor invasive metastasis and prognosis. Sema 5A of the study is ubiquitous in urine exosomes of people grouped at different ages, and is indifferently and stably expressed among groups.
Compared with the common clinical blood sample, the urine can be completely and noninvasively collected continuously in a large amount; can accumulate more kinds and larger amplitude of changes, and many pathophysiological changes of the organism can be reflected in urine. Some protein polypeptides with relatively small molecular weights such as hormones and cytokines are metabolized and excreted into urine soon after entering blood, and the probability of being detected in urine is greatly increased compared with that in blood; the possible protein degradation process in urine is completed before the sample is collected, and urine protein can be kept stable for a long period of time. The exosomes have membranous vesicle structures, the molecular stability is high, and the urine exosome proteomics research combines the advantages of the membranous vesicle structures and has the characteristics of good biomarker sources. Based on early-stage methodological fumbling, the experiment is expected to realize the application of noninvasive, convenient, quick and easily repeated urine exosome stable expression protein to urine internal reference markers, urine exosome research, clinical research of healthy people, related disease diagnosis, differential diagnosis, disease degree and activity judgment, treatment effect evaluation, monitoring, prognosis evaluation and mechanism research through urine protein or polypeptide research, and lays a foundation for the research of further urine polypeptide detection kits.
Disclosure of Invention
The invention aims to provide an application of a urine exosome nerve guiding molecule 5A and a polypeptide fragment thereof as urine internal reference markers and comparison markers in preparation of urine exosome detection, exosome research, clinical research of healthy people, related disease diagnosis, differential diagnosis, disease degree and activity judgment, treatment effect evaluation, monitoring, prognosis evaluation and mechanism research.
Preferably, the amino acid sequence of the urine exosomal nerve guidance molecule 5A comprises the amino acid sequence shown in SEQ ID No.1 (MKGTCVIAWL FSSLGLWRLA HPEAQGTTQC QRTEHPVISY KEIGPWLREF
RAKNAVDFSQ LTFDPGQKEL VVGARNYLFR LQLEDLSLIQ AVEWECDEAT
KKACYSKGKS KEECQNYIRV LLVGGDRLFT CGTNAFTPVC TNRSLSNLTE
IHDQISGMAR CPYSPQHNST ALLTAGGELY AATAMDFPGR DPAIYRSLGI
LPPLRTAQYN SKWLNEPNFV SSYDIGNFTY FFFRENAVEH DCGKTVFSRA
ARVCKNDIGG RFLLEDTWTT FMKARLNCSR PGEVPFYYNE LQSTFFLPEL
DLIYGIFTTN VNSIAASAVC VFNLSAIAQA FSGPFKYQEN SRSAWLPYPN
PNPHFQCGTV DQGLYVNLTE RNLQDAQKFI LMHEVVQPVT TVPSFMEDNS
RFSHVAVDVV QGREALVHII YLATDYGTIK KVRVPLNQTS SSCLLEEIEL
FPERRREPIR SLQILHSQSV LFVGLREHVV KIPLKRCQFY RTRSTCIGAQ
DPYCGWDVVM KKCTSLEESL SMTQWEQSIS ACPTRNLTVD GHFGVWSPWT
PCTHTDGSAV GSCLCRTRSC DSPAPQCGGW QCEGPGMEIA NCSRNGGWTP
WTSWSPCSTT CGIGFQVRQR SCSNPTPRHG GRVCVGQNRE ERYCNEHLLC
PPHMFWTGWG PWERCTAQCG GGIQARRRIC ENGPDCAGCN VEYQSCNTNP
CPELKKTTPW TPWTPVNISD NGGHYEQRFR YTCKARLADP NLLEVGRQRI
EMRYCSSDGT SGCSTDGLSG DFLRAGRYSA HTVNGAWSAW TSWSQCSRDC
SRGIRNRKRV CNNPEPKYGG MPCLGPSLEY QECNILPCPV DGVWSCWSPW
TKCSATCGGG HYMRTRSCSN PAPAYGGDIC LGLHTEEALC NTQPCPESWS
EWSDWSECEA SGVQVRARQC ILLFPMGSQC SGNTTESRPC VFDSNFIPEV
SVARSSSVEE KRCGEFNMFH MIAVGLSSSI LGCLLTLLVY TYCQRYQQQS
HDATVIHPVS PAPLNTSITN HINKLDKYDS VEAIKAFNKN NLILEERNKY
FNPHLTGKTY SNAYFTDLNN YDEY); 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 urine exosome nerve guiding molecule 5A and polypeptide fragment detection kit thereof.
Preferably, 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 an antibody fragment capable of specifically binding to the nerve directing molecule 5A and a polypeptide fragment thereof.
Preferably, the detection method comprises a mass spectrum method for directly detecting the nerve guidance molecule 5A and the polypeptide fragment 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 nerve guidance molecule 5A 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 a nerve guidance molecule 5A standard, a humanized tag antibody standard; preferably, the quality control product comprises: nerve guidance molecule 5A 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.
Firstly, collecting urine specimens of male and female healthy people of different ages, extracting urine exosomes by referring to a human urine proteome program (Human Urine Proteome Project, HKUPP) and European kidney and urine proteome (European Kidney and Urine Proteomics, euroKUP) group (http:// www.eurokup.org) standard operation procedures, identifying the urine exosomes by transmission electron microscopy, exosome nanoparticle tracking detection and western immunoblotting, constructing a normal human urine exosome protein map by adopting a Data independent scanning mode (Data-independent acquisition, DIA) by adopting a liquid-phase secondary mass spectrometry technology (liquid chromatography tandem mass spectrometry, LC-MS/MS), analyzing urine exosome stable indifferently expressed protein nerve guidance molecules 5A by using a bioinformatics analysis method, and verifying the urine exosome nerve guidance molecules 5A and the content of healthy human urine exosomes of different sexes of different ages by using a protein immunoblotting method.
According to the invention, research proves that Sema 5A and polypeptide fragments thereof of the urine exosomes of healthy people are stably expressed, so that the detection of the urine exosome nerve guiding molecules 5A and polypeptide fragments thereof can be applied to various purposes of urine internal reference markers, urine exosomes, healthy people and related disease 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 urine exosome nerve guiding molecule 5A 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 urinary exosomal nerve directing molecule 5A and polypeptide fragments thereof in healthy people.
FIG. 2 is a graph showing Western blotting results of urinary exosomal nerve guidance molecule 5A and polypeptide fragments thereof in healthy people.
Detailed Description
Example 1Urine specimen collection, exosome extraction preservation and identification
Collecting 100mL of clean middle-stage morning urine of 7 age groups of men and women covering children, teenagers, middle-aged and elderly people in normal life state. The group-entering personnel have no hypertension, diabetes, cardiovascular and cerebrovascular diseases and other system diseases, liver and kidney dysfunction, no medicine taking history in the last 1 month and apparent health. Taking 100mL of clean mid-morning urine of the crowd in a normal life state after the agreement of the parties. Exosomes in concentrated urine samples were extracted, resuspended in pre-chilled PBS and stored in a-80℃refrigerator, with reference to standard procedures for HKUPP and EuroKUP (http:// www.eurokup.org). The urine exosomes are identified by transmission electron microscopy, exosome nanoparticle tracking detection and western blotting.
Example 2Urine exosome polypeptide mass spectrometry
And (3) carrying out protein extraction and enzymolysis desalination on the urine sample, and measuring the concentration of the extracted protein. By advanced liquid-phase secondary mass spectrometry (LC-MS/MS), a data independent scanning mode (DIA) is adopted to construct urine exosome protein maps of normal people with different ages and different sexes. All experiments were performed in triplicate and all values are expressed as mean ± standard deviation. Fold change 1.5 times, the analysis result of the differential protein was obtained, and FDR <0.05 was considered statistically significant.
Example 3Bioinformatics analysis
The homosapiens database was used. The mass spectrum original file was analyzed by PD software, homology classification was performed using database ortholog cluster (Clusters of Orthologous Groups, COG), statistical analysis was performed on cell components (cellular component, CC), molecular functions (molecular function, MF), biological processes (biological process, BP) using Gene Ontology (GO) using protein database, and stable expression proteins were screened.
Example 4Verification of urinary exosomatic nerve-directing molecule 5A
The content of the urine exosome nerve guiding molecule 5A in the healthy population is shown in figure 1 by mass spectrometry and the result is shown in figure 2 by using western blotting to detect the nerve guiding molecule 5A in the urine exosome of the healthy population. The nerve guidance molecule 5A is indifferently and stably expressed in urine of healthy people.
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> application of nerve guidance molecule 5A and polypeptide fragment thereof as urine reference marker
<130> 21SEMA5A-CN
<140> 21SEMA5A-CN
<141> 2021-11-30
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1074
<212> PRT
<213> Human Urine
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Met Lys Gly Thr Cys Val Ile Ala Trp Leu Phe Ser Ser Leu Gly Leu
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Trp Arg Leu Ala His Pro Glu Ala Gln Gly Thr Thr Gln Cys Gln Arg
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Thr Glu His Pro Val Ile Ser Tyr Lys Glu Ile Gly Pro Trp Leu Arg
35 40 45
Glu Phe Arg Ala Lys Asn Ala Val Asp Phe Ser Gln Leu Thr Phe Asp
50 55 60
Pro Gly Gln Lys Glu Leu Val Val Gly Ala Arg Asn Tyr Leu Phe Arg
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Leu Gln Leu Glu Asp Leu Ser Leu Ile Gln Ala Val Glu Trp Glu Cys
85 90 95
Asp Glu Ala Thr Lys Lys Ala Cys Tyr Ser Lys Gly Lys Ser Lys Glu
100 105 110
Glu Cys Gln Asn Tyr Ile Arg Val Leu Leu Val Gly Gly Asp Arg Leu
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Phe Thr Cys Gly Thr Asn Ala Phe Thr Pro Val Cys Thr Asn Arg Ser
130 135 140
Leu Ser Asn Leu Thr Glu Ile His Asp Gln Ile Ser Gly Met Ala Arg
145 150 155 160
Cys Pro Tyr Ser Pro Gln His Asn Ser Thr Ala Leu Leu Thr Ala Gly
165 170 175
Gly Glu Leu Tyr Ala Ala Thr Ala Met Asp Phe Pro Gly Arg Asp Pro
180 185 190
Ala Ile Tyr Arg Ser Leu Gly Ile Leu Pro Pro Leu Arg Thr Ala Gln
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Tyr Asn Ser Lys Trp Leu Asn Glu Pro Asn Phe Val Ser Ser Tyr Asp
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Ile Gly Asn Phe Thr Tyr Phe Phe Phe Arg Glu Asn Ala Val Glu His
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Asp Cys Gly Lys Thr Val Phe Ser Arg Ala Ala Arg Val Cys Lys Asn
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Asp Ile Gly Gly Arg Phe Leu Leu Glu Asp Thr Trp Thr Thr Phe Met
260 265 270
Lys Ala Arg Leu Asn Cys Ser Arg Pro Gly Glu Val Pro Phe Tyr Tyr
275 280 285
Asn Glu Leu Gln Ser Thr Phe Phe Leu Pro Glu Leu Asp Leu Ile Tyr
290 295 300
Gly Ile Phe Thr Thr Asn Val Asn Ser Ile Ala Ala Ser Ala Val Cys
305 310 315 320
Val Phe Asn Leu Ser Ala Ile Ala Gln Ala Phe Ser Gly Pro Phe Lys
325 330 335
Tyr Gln Glu Asn Ser Arg Ser Ala Trp Leu Pro Tyr Pro Asn Pro Asn
340 345 350
Pro His Phe Gln Cys Gly Thr Val Asp Gln Gly Leu Tyr Val Asn Leu
355 360 365
Thr Glu Arg Asn Leu Gln Asp Ala Gln Lys Phe Ile Leu Met His Glu
370 375 380
Val Val Gln Pro Val Thr Thr Val Pro Ser Phe Met Glu Asp Asn Ser
385 390 395 400
Arg Phe Ser His Val Ala Val Asp Val Val Gln Gly Arg Glu Ala Leu
405 410 415
Val His Ile Ile Tyr Leu Ala Thr Asp Tyr Gly Thr Ile Lys Lys Val
420 425 430
Arg Val Pro Leu Asn Gln Thr Ser Ser Ser Cys Leu Leu Glu Glu Ile
435 440 445
Glu Leu Phe Pro Glu Arg Arg Arg Glu Pro Ile Arg Ser Leu Gln Ile
450 455 460
Leu His Ser Gln Ser Val Leu Phe Val Gly Leu Arg Glu His Val Val
465 470 475 480
Lys Ile Pro Leu Lys Arg Cys Gln Phe Tyr Arg Thr Arg Ser Thr Cys
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Ile Gly Ala Gln Asp Pro Tyr Cys Gly Trp Asp Val Val Met Lys Lys
500 505 510
Cys Thr Ser Leu Glu Glu Ser Leu Ser Met Thr Gln Trp Glu Gln Ser
515 520 525
Ile Ser Ala Cys Pro Thr Arg Asn Leu Thr Val Asp Gly His Phe Gly
530 535 540
Val Trp Ser Pro Trp Thr Pro Cys Thr His Thr Asp Gly Ser Ala Val
545 550 555 560
Gly Ser Cys Leu Cys Arg Thr Arg Ser Cys Asp Ser Pro Ala Pro Gln
565 570 575
Cys Gly Gly Trp Gln Cys Glu Gly Pro Gly Met Glu Ile Ala Asn Cys
580 585 590
Ser Arg Asn Gly Gly Trp Thr Pro Trp Thr Ser Trp Ser Pro Cys Ser
595 600 605
Thr Thr Cys Gly Ile Gly Phe Gln Val Arg Gln Arg Ser Cys Ser Asn
610 615 620
Pro Thr Pro Arg His Gly Gly Arg Val Cys Val Gly Gln Asn Arg Glu
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Glu Arg Tyr Cys Asn Glu His Leu Leu Cys Pro Pro His Met Phe Trp
645 650 655
Thr Gly Trp Gly Pro Trp Glu Arg Cys Thr Ala Gln Cys Gly Gly Gly
660 665 670
Ile Gln Ala Arg Arg Arg Ile Cys Glu Asn Gly Pro Asp Cys Ala Gly
675 680 685
Cys Asn Val Glu Tyr Gln Ser Cys Asn Thr Asn Pro Cys Pro Glu Leu
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Lys Lys Thr Thr Pro Trp Thr Pro Trp Thr Pro Val Asn Ile Ser Asp
705 710 715 720
Asn Gly Gly His Tyr Glu Gln Arg Phe Arg Tyr Thr Cys Lys Ala Arg
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Leu Ala Asp Pro Asn Leu Leu Glu Val Gly Arg Gln Arg Ile Glu Met
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Arg Tyr Cys Ser Ser Asp Gly Thr Ser Gly Cys Ser Thr Asp Gly Leu
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Ser Gly Asp Phe Leu Arg Ala Gly Arg Tyr Ser Ala His Thr Val Asn
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Gly Ala Trp Ser Ala Trp Thr Ser Trp Ser Gln Cys Ser Arg Asp Cys
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Ser Arg Gly Ile Arg Asn Arg Lys Arg Val Cys Asn Asn Pro Glu Pro
805 810 815
Lys Tyr Gly Gly Met Pro Cys Leu Gly Pro Ser Leu Glu Tyr Gln Glu
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Cys Asn Ile Leu Pro Cys Pro Val Asp Gly Val Trp Ser Cys Trp Ser
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Pro Trp Thr Lys Cys Ser Ala Thr Cys Gly Gly Gly His Tyr Met Arg
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Lys Arg Cys Gly Glu Phe Asn Met Phe His Met Ile Ala Val Gly Leu
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Ser Ser Ser Ile Leu Gly Cys Leu Leu Thr Leu Leu Val Tyr Thr Tyr
980 985 990
Cys Gln Arg Tyr Gln Gln Gln Ser His Asp Ala Thr Val Ile His Pro
995 1000 1005
Val Ser Pro Ala Pro Leu Asn Thr Ser Ile Thr Asn His Ile Asn Lys
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Leu Asp Lys Tyr Asp Ser Val Glu Ala Ile Lys Ala Phe Asn Lys Asn
1025 1030 1035 1040
Asn Leu Ile Leu Glu Glu Arg Asn Lys Tyr Phe Asn Pro His Leu Thr
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Gly Lys Thr Tyr Ser Asn Ala Tyr Phe Thr Asp Leu Asn Asn Tyr Asp
1060 1065 1070
Glu Tyr

Claims (9)

1. The application of the nerve guidance molecule 5A and the polypeptide fragment thereof as urine exosome stable expression protein serving as urine internal reference markers and comparison markers in preparation of urine exosome detection, exosome research, health crowd clinical research, related disease diagnosis, differential diagnosis, disease degree and activity judgment, treatment effect evaluation, monitoring, prognosis evaluation and mechanism research.
2. The use according to claim 1, wherein the amino acid sequence of the urinary exosomal nerve guidance molecule 5A comprises the amino acid sequence 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 urine exosomal nerve guidance molecule 5A and a polypeptide fragment detection kit 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 an antibody fragment capable of specifically binding to the nerve guidance molecule 5A and a polypeptide fragment thereof.
5. The use according to claim 3, wherein the detection method comprises a method for directly detecting mass spectra of the nerve-directing molecule 5A and its polypeptide fragments, and the like, and a kit thereof.
6. The use according to claim 3, wherein the detection method comprises a method for directly detecting the nerve-directing molecule 5A and its polypeptide fragment or its related nucleic acid detection and its related kit.
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 of claim 7, wherein the standard comprises a nerve-directing molecule 5A standard, a humanized tag antibody standard; preferably, the quality control product comprises: nerve guidance molecule 5A control, humanized tag antibody 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.
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.
CN202111561840.1A 2021-12-20 2021-12-20 Application of nerve guidance molecule 5A and polypeptide fragment thereof as urine reference marker Pending CN117147850A (en)

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