CN108267578A - The detection method of serum IgG core fucosylation level - Google Patents

The detection method of serum IgG core fucosylation level Download PDF

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CN108267578A
CN108267578A CN201711475309.6A CN201711475309A CN108267578A CN 108267578 A CN108267578 A CN 108267578A CN 201711475309 A CN201711475309 A CN 201711475309A CN 108267578 A CN108267578 A CN 108267578A
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serum igg
igg
core
core fucosylation
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李文哲
李志�
李明
梁伟
李雪滢
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Dalian Medical University
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Abstract

The present invention disclose a kind of serum IgG core fucosylation level detection method, this method including the use of in agglutinin specific recognition serum IgG the step of core fucosido, not including being sampled from organism the step of, the blood serum sample is vitro samples.The agglutinin is the aspergillus oryzae agglutinin AOL albumen (structural domain) of specific recognition core fucose base, is obtained by prokaryotic expression, and amino acid sequence is as shown in SEQ ID NO.1；As FleA genetic fragment coding of the sequence as shown in SEQ ID NO.2.This method combines clinical antinuclear antibodies titre detection, and the detection direction of screening, autoimmune disease in autoimmune disease has potential application value, potential treatment means are provided for possible immunoregulation intervention.

Description

The detection method of serum IgG core fucosylation level
Technical field
The invention belongs to DNA techniques fields, are related to the detection of glycoprotein, and in particular to serum IgG core fucose base The detection of change.
Background technology
Autoimmune disease (autoimmune diseases) refers to that body occurs immune response to autoantigen and leads Cause the disease caused by damaged self tissue.Have the characteristics that following：1) patient has apparent family's tendentiousness；2) in blood There are high titre autoantibodies；3) process of recurrent exerbation and chronic delay is often presented in autoimmunity disease；4) autoimmunity is after being ill Phase is often with the dysfunction and complication of the vitals such as liver, kidney, intestines.It there is no radical cure method can only anti symptom treatment at present.Such as Fruit can early detection autoimmune disease and monitor its development and change, in time treatment, the generation of complication can be delayed.
At present, the screening means of autoimmune disease predominantly detect antinuclear antibodies (ANA), rheumatoid factor (RF), resist Double-stranded DNA (ds-DNA) antibody resists nonhistones (ENA) polypeptide spectrum.However, their sensibility and specificity is not generally high. Therefore, a kind of broad covered area, sensibility height, good, simple operation the hematology method of specificity are found out and is used for autoimmune disease The early screening of disease, has great importance at prognosis evaluation.In terms of age incidence, incidence rises apparent after 50 years old.Cause This, screening was carried out since 40 years old for autoimmune disease, had very big social effect.
Sugar chain almost takes part in all life processes, the change of sugar chain structure and disease as a kind of biological information molecule Generation, the development of the especially major diseases such as autoimmune disease, tumour, infection are closely related.IgG is that content is most in serum More immunoglobulin classes (account for human serum Ig 80%), crystallized region (Fc) contain the glycosylation modified conservative positions of unique N- Point, i.e. Asn297Sugar chain is since the terminal saccharide connected on seven sugar of core is different (Fig. 1), and there is kind, tissue, age and property Not specificity.IgG glycosylations are closely related with function.It is connected to Asn297Sugar chain can maintain the quaternary structure and Fc of antibody The thermal stability of section, and pass through influence IgG molecules and FcRs, C1q and FcRn with reference to and adjust the antibody of IgG molecules respectively Dependent cells toxic action (ADCC), the cytotoxicity (CDC) of Complement Dependent and half-life period.The Fc sections sugar of research prompting IgG Base decorating state and a variety of diseases are closely related, such as generation, development with rheumatoid arthritis, the low galactolipins of IgG The degree of baseization modification increases；The ConA that advanced ovarian cancer patient possesses high-level asymmetric oligosaccharide structure combines (high mannose) IgG.
In the processing and identification process of glycoprotein, the core fucosylation on N- connection glycoprotein plays very important Effect.Core fucosyltransferase (Fut8) is unique glycosyl transferase (Fig. 2) of catalytic core fucosylation modification. Core fucose appears in the different phase of N- connection glycan, and can influence the conformation and adaptability of 2 antenna oligosaccharides of N- connections. We are previous the study found that core on Transforming growth factor-β1 (TGF β -1) receptor and/or epidermal growth factor (EGF) receptor After heart fucosylation missing, show significantly to activate exception, be attributed to reduction of the ligand to receptor affinity.Also report Road removes IgG1-CHAfter core fucose on 2 regions, ADCC can enhance 50-100 times.To sum up, the core of Fut8 mediations Heart fucosylation is extremely important to protein translation post-processing, conformation, stability and the function of regulation protein.
At present, related article and patent are as follows：
1. title：IgG core fucosylations level detection method and its application；Publication number：CN102175839A.
IgG core fucosylations level is mainly detected by LCA gel chromatographies, and applied to diagnosing hepatic diseases.
2. title：For the product of the screening of malignant tumour correlation and assessment, application and method；Publication number： CN105277718A。
Mainly changed by detecting immunoglobulin G surface double antenna complexity N sugar-chain ends galactolipin in blood to disliking Property tumour carry out screening, early diagnosis and prognosis evaluation etc..
3. title：Alpha-fetoprotein variant separating kit and its composition reagent and application for diagnosing cancer of liver；It is open Number：CN102879567A.
A kind of alpha-fetoprotein variant separating kit for diagnosing cancer of liver.
Invention content
The invention belongs to protein and sugar to learn technical field, is related to the detection of glycoprotein, and in particular to serum IgG core rock Application of the algae level of glycosylation detection in autoimmunity disease diagnosis, is detected including IgG core fucosylations level.This method It is detected with reference to clinical antinuclear antibodies titre, there is good application prospect in the screening of autoimmune disease.
1. the object of the present invention is to provide a kind of serum IgG core fucosylation level detection method, this method is certainly The detection direction of body immunity disease has potential application value, such as：The inspection of patients serum IgG core fucosylation levels It surveys, potential treatment means is provided for possible immunoregulation intervention.
This method is including the use of the step of core fucosido, this method is not wrapped in agglutinin specific recognition serum IgG Include the infectivity of detection blood serum sample；And the method include from organism sample the step of, the blood serum sample is in vitro Sample.This method does not include the molecule of other in addition to IgG of detection blood serum sample.Due to what is used selected by the present invention Protein G (Acrobiosystems, RPG-S3140) and AOL are that prokaryotic expression is obtained, and Protein G and AOL do not have It is glycosylation modified.Therefore, the various molecules for participating in the detection method only have serum IgG to be checked to have core fucose based structures.
In the case of preferred, the agglutinin is the aspergillus oryzae agglutinin AOL eggs of specific recognition core fucose base (structural domain) in vain, is obtained by prokaryotic expression.
In the case of preferred, the amino acid sequence of the agglutinin AOL albumen is as shown in SEQ ID NO.1；By sequence FleA genetic fragments coding as shown in SEQ ID NO.2.
In the case of preferred, the method for core fucosido in the specific recognition serum IgG using agglutinin.
The step of including A or B：
Core fucosylation is horizontal in A agglutinins immunoblot method detection serum IgG；
B detects core fucosylation level in serum IgG sample by sandwich method ELISA method.
In the case of preferred, the side of core fucosylation in sandwich method ELISA method detection serum IgG sample Method is that coating recombinates protein G structural domains or streptococcus pneumoniae polypeptides (190-384), is incubated blood to be checked successively again first Final proof product, reaction biotin-AOL (AOL as described in claim 1 of biotin labeling), HRP-avidin (horseradish peroxidating The avidin of object enzyme label), it is eventually adding reaction substrate.The system includes the exploitation of sandwich method ELISA and microplate reader (Thermo Company), wherein microplate reader can analyze proteinG/IgG/biotin-AOL/avidin-HRP levels in reaction solution.Folder Heart method ELISA method is efficient, special, suitable for the screening of clinical autoimmunity disease.
In the case of preferred, recombination protein G used in the present invention are Acrobiosystems, RPG-S3140, reality On border, the albumen specifically combined with human IgG can be used, such as：Streptococcus pneumoniae polypeptides (190-384).
In the case of preferred, the side of core fucosylation level in agglutinin immunoblot method detection serum IgG Method be sequentially add blood serum sample SDS-PAGE electrophoresis to be checked, go to pvdf membrane, reaction biotin labeling such as claim 1 The avidin of AOL, HRP (horseradish peroxidase) label, last ECL colour developings.
In the case of preferred, this method further includes step：
The expression quantity of IgG in C Western blot immunoblot methods detection serum；
D detects serum IgG expression quantity and the horizontal ratio of core fucose base.
In the case of preferred, the method for the expression quantity of IgG includes step in western blot method detection serum：Successively Add in blood serum sample SDS-PAGE electrophoresis to be checked, go to pvdf membrane, donkey that reaction HRP (horseradish peroxidase) is marked it is anti-human IgG antibody, last ECL colour developings.
In the case of preferred, the serum IgG core fucose base level and the method for IgG expression quantity ratios, the party Method further comprises that calculate serum IgG core fucose base level examines with IgG expression quantity ratio (ratio of AOL/IgG total amounts) It surveys.In this way, it can directly judge whether the patient has the tendency that autoimmune disease.Its by Image Lab with 6 softwares of Graphpad Prism carry out significant difference data analysis.
Furthermore, it is possible to method as discussed above according to the present invention, utilizes agglutinin described above, i.e. specific recognition core The aspergillus oryzae agglutinin AOL albumen (structural domain) of heart fucosido is prepared a kind of horizontal for serum IgG core fucosylation Detection kit.
Description of the drawings
IgG molecules are a kind of glycoprotein of core fucosylation in Fig. 1 serum；
Fig. 2 core fucosyltransferases are catalyzed response diagram；
The prokaryotic expression of Fig. 3 biotin labeling AOL structural domains；
Fig. 4 patient's IgG measurement results；
Fig. 5 patient's AOL measurement results；
Fig. 6 patient's AOL/IgG ratio statistical analysis；
Fig. 7 patient's antinuclear antibodies titre statistical analysis；
Fig. 8 sandwich method ELISAs method detects autoimmune disease patient's serum.
Specific embodiment
Following nonlimiting examples can make those of ordinary skill in the art be more fully understood the present invention, but not with Any mode limits the present invention.
Raw material, reagent or biomaterial used in the embodiment of the present invention, unless otherwise specified, can be by conventional method It prepares or is obtained by commercial sources.
Gel imager (Bio-Rad) can analyze IgG core fucosylation levels.
Embodiment 1
The detection and analysis of Normal group and autoimmune disease group IgG core fucosylation levels
1. sample collection
One 2016 Nian9Yue Lai The Central Hospital of Dalian City of September in 2015 is collected to receive to treat and clarify a diagnosis as autoimmunity Property disease serum sample 37, wherein man 11, female 26, average age 51.6 ± 13.2 years old.In addition health examination is selected Person 20 is as Normal group, average age 51.2 ± 10.3 years old.Empty stomach whole blood is extracted, stands 30 minutes, 3000rpm, l0min.Upper serum is drawn, -80 DEG C of refrigerators save backup.Antinuclear antibodies titre (table 1 in autoimmune disease patient's serum It is shown).
1 autoimmunity disease patients serum's sample of table
2 prokaryotic expressions specifically bind the AOL agglutinins of core fucose, for follow-up clinical sample core fucose base The detection of change
Prepared by prokaryotic expression AOL, be as follows：
A. to extract obtained aspergillus oryzae RNA as template, reverse transcription obtains cDNA, then expands FleA genetic fragments (Encoded Lectin AOL albumen).The amino acid sequence of the agglutinin AOL albumen is as shown in SEQ ID NO.1；By sequence such as FleA genetic fragments coding shown in SEQ ID NO.2.Sense primer (5'-CCGGAATTCA TGTCTACTCCTGGCGCCCAA-3') including EcoR I restriction enzyme sites, downstream primer (5'-CCGCTCGAGCTACTCAGC CGG AGGGAGAGC-3') including Xho I restriction enzyme sites.PCR product is attached with pMD19-T carriers, Transformed E coli TOP10 senses It by state cell, in 37 DEG C of overnight incubations of tablet of Amp resistances, is sequenced after picking single bacterium colony culture, builds pMD19-T- FleA plasmids.
B. correct pMD19-T-FleA and pET-28a (+) will be sequenced and be carried out at the same time EcoR I and Xho I double digestions, return It is attached after receipts, connection product is converted into e. coli bl21 competent cell, then structure recombinant expression plasmid send company Sequencing.
Correct bacterium solution will be sequenced and continue culture, induced expression recombinant protein, i.e.,：1mL is taken to be inoculated into the training of 50mL LB liquid It supports in base, 37 DEG C, is cultivated 30 minutes 1 hour under the conditions of 200r/min, add in 0.5mM derivants IPTG (Isopropyl β-D-1- Thiogalactoside), 0,2,4 and 5h is cultivated respectively, induces the expression of FleA recombinant proteins, SDS-PAGE detections.
C. it will contain prokaryotic expression carrier, the expression bacterium E.coli BL21 of pET20b-AOL are in 37 DEG C of shake cultures (170rpm)；Expand culture, same procedure induced expression, 6500rmp centrifugations after 1% ratio of recombinant bacterium inoculation after expression 10min collects cell, suspends, and Ultrasonic Cell Disruptor smudge cells (20% power crushes 10min) are detected through SDS-PAGE.Finally Pass through AKATA instrument nickel ion affinity chromatograph column purification albumen.
Testing result is：
It is connect by the FleA genetic fragments of PCR amplification and purifying with pMD19-T carriers, is built into pMD19-T-FleA matter Grain after the processing of EcoR I and Xho I restriction enzymes double zyme cuttings, detects pET-28a (+) carrier (5369bp) and FleA Genetic fragment (935bp) (Fig. 3 A).After IPTG induces 0,2 and 4h, AOL albumen (molecular weight about 34kDa) (Fig. 3 B) is given expression to.Expand The purity of AOL albumen that SDS-PAGE is detected after big culture is close to 85% (Fig. 3 C).Therefore, destination protein success table is as a result proved It reaches, by the way that subsequent experimental can be carried out after purification.
The agglutinin AOL of 3 pairs of prokaryotic expressions carries out biotin labeling；
A. LCB-NHS (long-chain biological element NHS esters) solution is prepared with DMSO (6mg/ml, 6.5mM)；
B. 10ul LCB-NHS are added in 1ml 2mg/ml antibody, and be incubated at room temperature 50 minutes；
C. 0.5ml 1M Tris-HCl (pH 8.0) are added in and terminate reaction；
D. it is dialysed 24-48 hours with 1X PBS；
E. the NaN of 0.1% (w/v) is added in3；
4Western immunoblot methods detect autoimmune disease group IgG and Normal group IgG in blood serum sample respectively Expression；
A. it after blood serum sample is by SDS-PAGE electrophoresis, goes on pvdf membrane, with 5% bovine serum albumin(BSA) (BSA) room temperature Close 1h；
B. incubation at room temperature HRP label donkeys anti-human IgG antibodies (1:10000) 1 hour；
C.TBST is washed, and 10 minutes every time, is washed 5 times；
D. ECL reaction solution (A liquid, B liquid 1 are added in:1 mixing)；
E. it is developed the color by gel imager, detects protein band；
Testing result is：
Blood serum sample after dilution detects IgG in serum after SDS-PAGE electrophoresis, with donkey anti-human IgG antibodies and contains Amount.It was found that compared with Normal group, autoimmune disease group IgG content significantly increases, and difference is respectively provided with statistics meaning Justice (P<0.05) (shown in Fig. 4).Show that IgG content is higher than normal level in autoimmune disease patient.
5 agglutinin immunoblot methods detect respectively in serum IgG autoimmune disease group core fucosylation with it is normal Control group core fucosylation；
A. it after blood serum sample is by SDS-PAGE electrophoresis, goes on pvdf membrane, is closed with 5% bovine serum albumin(BSA) (BSA) Afterwards, it is incubated biotin labeling AOL (1:10000-1:20000) 4 night is spent；
B.TBST is washed, and 10 minutes every time, is washed 5 times；
C. reaction HRP labels Avidin (1:10000) room temperature 1 hour
D.TBST is washed, and 10 minutes every time, is washed 5 times；
E. ECL reaction solution (A liquid, B liquid 1 are added in:1 mixing)；
F. it is developed the color by gel imager, detects protein band；
Testing result is：
The significantly raising, and difference has compared with Normal group of autoimmune disease group core fucosylation level Statistically significant (P<0.001) (shown in Fig. 5).
The AOL/IgG average values of Normal group are 0.58, and the AOL/IgG average values of autoimmune disease group are 1.09, and difference is respectively provided with statistical significance (P<0.05), there is certain reference detection specificity and sensitivity (Fig. 6 institutes Show).
High-caliber antinuclear antibodies (ANA) is able to detect that in SLE patients serums.Compared with normal serum sample, patient (antinuclear antibodies titre is 1 to serum:100-1:1000) protein core fucosylation level significantly reduces, and difference is respectively provided with statistics Learn meaning (P<0.05)；Compared with normal serum sample, (antinuclear antibodies titre is 1 to patients serum:1000-1:3200) pyrenoids Heart fucosylation level significantly reduces, and difference is respectively provided with statistical significance (P<0.01) (shown in Fig. 7).
May have these results show high-level core fucosylation to SLE severity and pathogenesis certain Effect.
Embodiment 2
Sandwich method ELISA method detects autoimmune disease patient's serum
1 coating
With the carbonate buffer solution dilution coating protein G of pH 9.6, concentration is made to reach 5-20 μ g/ml, per 50 μ l of hole Coated elisa plate, 4 DEG C of coatings are overnight.
2 board-washings
Coating buffer is dried, is washed with PBST, 200 μ l/ holes, board-washing liquid is dried after each 30s, in triplicate.
3 closings
Add in 200 μ l/ holes of confining liquid, 37 DEG C of wet box warm bath 30min.
4 add in tested blood serum sample
Add in 1：10-1：100 times of diluted normal human sera samples and patient serum sample.37 DEG C of reaction 1h.It is negative right According to for PBS；Positive control is patient diagnosed's blood serum sample.
5 board-washings
It is washed with PBST, 200 μ l/ holes, board-washing liquid is dried after each 30s, in triplicate.
6 primary antibodies react
Add in 1：1000-1：The biotin labeling of 20000 doubling dilutions AOL (obtained by prokaryotic expression, and with biology Element is marked) 100 μ l/ holes, 37 DEG C of wet box warm bath 1h.
7 board-washings
Primary antibody is dried, is washed with PBST, 200 μ l/ holes, board-washing liquid is dried after each 30s, in triplicate.
8 secondary antibodies react
Avidin labels HRP is done 1:5000-1:10000 dilutions, 100 μ l/ holes add in ELISA Plate, 37 DEG C of wet box warm bath 1h。
9 board-washings
Secondary antibody is dried, is washed with PBST, 200 μ l/ holes, board-washing liquid is dried after each 30s, in triplicate.
10 substrates
100 μ l/ holes of o-phenylenediamine (OPD) liquid are added in, 37 DEG C are protected from light 15min, and microplate reader measures OD492nmValue.
Testing result is：
Sandwich method ELISA result shows that autoimmune disease patient's IgG core fucosylation levels are significantly higher than just Ordinary person, such as Fig. 8.Detectable core fucosylation IgG concentration ranges are 0.01~1000pmol/L, and detection lower limit is 0.01 ~0.05pmol/L.The present invention substantially increases the specificity and sensibility of detection autoimmune disease, easy to operate, repetition Property high, the expression quantity of core fucosylation IgG in present invention detection patients serum, the screening for autoimmune disease provides Experimental data.
Sequence table
<110>Dalian Medical Univ
<120>The detection method of serum IgG core fucosylation level
<130> 2014
<160> 2
<170> PatentIn version 3.3
<210> 1
<211> 311
<212> Protein
<213>The amino acid sequence of agglutinin AOL albumen
<400> 1
MSTPGAQEVL FRTGIAAVNS TNHLRVYFQD SHGSIRESLY ESGWANGTAK NVIAKAKLGT 60
PLAATSKELK NIRVYSLTED NVLQEAAYDS GSGWYNGALA GAKFTVAPYS RIGSVFLAGT 120
NALQLRIYAQ KTDNTIQEYM WNGDGWKEGT NLGVALPGTG IGVTCWRYTD YDGPSIRVWF 180
QTDNLKLVQR AYDPHTGWYK ELTTIFDKAP PRCAIAATNF NPGKSSIYMR IYFVNSDNTI 240
WQVCWDHGQG YHDKRTITPV IQGSEIAIIS WEGPELRLYF QNGTYVSAIS EWTWGKAHGS 300
QLGRRALPPA E 311
<210> 2
<211> 936
<212> DNA
<213>The nucleotide sequence of FleA genetic fragments
<400> 2
atgtctactc ctggcgccca agaagttctt ttccgtaccg gaattgctgc ggtgaactcc 60
accaaccatc tccgtgtcta ctttcaggac tctcacggta gtattcgcga gagtctctat 120
gagagtgggt gggcgaatgg tacggcaaag aatgttatcg ccaaggcgaa gcttggtacc 180
cctctcgctg cgacatctaa ggaactgaaa aatattcgtg tctacagtct taccgaagac 240
aacgtccttc aggaagctgc ttatgatagt ggcagcggat ggtacaacgg cgcgctggct 300
ggcgctaaat tcacagttgc tccttactct cgaatcgggt ctgtctttct ggcaggaacg 360
aatgcgttgc agttgcgtat ctatgcccag aaaactgata acacgataca ggagtatatg 420
tggaatggag acggctggaa ggaaggcaca aaccttggag ttgcgcttcc tggcactggt 480
attggagtta cttgctggcg ctacaccgat tacgatggtc caagcattag ggtctggttc 540
caaaccgaca atttgaagct tgtccagcga gcatatgatc cccataccgg atggtataag 600
gaactgacta ccatctttga caaggctcct cctcgctgtg caatcgcagc cacgaacttt 660
aaccccggta aaagtagcat ttacatgcgg atctattttg tcaactctga caacacaatt 720
tggcaagtgt gttgggatca tggccaagga taccatgaca agagaaccat tacaccagtc 780
attcagggct cggaaattgc gatcattagc tgggaagggc ctgagctgcg tctgtacttt 840
caaaatggca catatgtcag tgccattagt gagtggacat ggggcaaagc acacggatcg 900
cagctgggtc gccgggctct ccctccggct gagtag 936

Claims (10)

  1. A kind of 1. method for detecting serum IgG core fucosylation level, which is characterized in that this method is including the use of agglutinin In specific recognition serum IgG the step of core fucosido, the method include from organism sample the step of, the blood Final proof product are vitro samples.
  2. 2. the method for detection serum IgG core fucosylation level according to claim 1, the agglutinin are special The aspergillus oryzae agglutinin AOL albumen of opposite sex identification core fucose base, is obtained by prokaryotic expression.
  3. 3. the method for detection serum IgG core fucosylation level according to claim 2, which is characterized in that described Agglutinin AOL albumen amino acid sequence as shown in SEQ ID NO.1；As FleA base of the sequence as shown in SEQ ID NO.2 Because of fragment coding.
  4. 4. the method for detection serum IgG core fucosylation level according to claim 3, which is characterized in that described The method using core fucosido in agglutinin specific recognition serum IgG, the step of including A or B：
    Core fucosylation is horizontal in A agglutinins immunoblot method detection serum IgG；
    B detects core fucosylation level in serum IgG sample by sandwich method ELISA method.
  5. 5. the method for detection serum IgG core fucosylation level according to claim 4, which is characterized in that described Sandwich method ELISA method detection serum IgG sample in the method for core fucosylation be coated with first protein G or Streptococcus pneumoniae polypeptides (190-384), be incubated successively again blood serum sample to be checked, reaction biotin labeling such as claim 1 The avidin of AOL, HRP (horseradish peroxidase) label, is eventually adding reaction substrate.
  6. 6. the method for detection serum IgG core fucosylation level according to claim 5, the protein G It is streptococcus pneumoniae polypeptides (190-384).
  7. 7. the method for detection serum IgG core fucosylation level according to claim 4, the agglutinin trace In method detection serum IgG the method for core fucosylation level be sequentially add blood serum sample SDS-PAGE electrophoresis to be checked, Go to pvdf membrane, AOL, HRP as described in claim 1 (horseradish peroxidase) of reaction biotin labeling are marked Avidin, last ECL colour developings.
  8. 8. the method for detection serum IgG core fucosylation level according to claim 1, which is characterized in that the party Method further includes step：
    The expression quantity of IgG in C western blots method detection serum；
    D detects serum IgG expression quantity and the horizontal ratio of core fucose base.
  9. 9. the method for detection serum IgG core fucosylation level according to claim 8, which is characterized in that The method of the expression quantity of IgG includes step in western blot method detection serum：Sequentially add blood serum sample SDS- to be checked PAGE electrophoresis goes to pvdf membrane, the donkey anti-human IgG antibodies that reaction HRP (horseradish peroxidase) is marked, last ECL colour developings.
  10. 10. the method for detection serum IgG core fucosylation level according to claim 8, which is characterized in that described Serum IgG core fucose base level and IgG expression quantity ratios method, be by Image Lab and Graphpad 6 softwares of Prism carry out significant difference analysis.
CN201711475309.6A 2017-12-29 2017-12-29 The detection method of serum IgG core fucosylation level Pending CN108267578A (en)

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Publication number Priority date Publication date Assignee Title
CN110031632A (en) * 2019-04-03 2019-07-19 中国医学科学院北京协和医院 A kind of biomarker of Disease Activity and application thereof that reflection IgG4 is diseases related
CN110045127A (en) * 2019-04-03 2019-07-23 中国医学科学院北京协和医院 A kind of biomarker and application thereof of the diseases related multiple organ involvement of IgG4
CN110045126A (en) * 2019-04-03 2019-07-23 中国医学科学院北京协和医院 A kind of biomarker and application thereof for diagnosis of autoimmune pancreatitis
CN110045125A (en) * 2019-04-03 2019-07-23 中国医学科学院北京协和医院 A kind of biomarker and application thereof for diagnosing retroperitoneal fibrosis
CN110045126B (en) * 2019-04-03 2022-08-09 中国医学科学院北京协和医院 Biomarker for diagnosing autoimmune pancreatitis and application thereof
CN110045125B (en) * 2019-04-03 2022-08-09 中国医学科学院北京协和医院 Biomarker for diagnosing retroperitoneal fibrosis and application thereof

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Application publication date: 20180710