CN110467665A - The specific antigen epitope peptide of Occludin 31kDa degradation fragment and its application - Google Patents
The specific antigen epitope peptide of Occludin 31kDa degradation fragment and its application Download PDFInfo
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- CN110467665A CN110467665A CN201910842179.8A CN201910842179A CN110467665A CN 110467665 A CN110467665 A CN 110467665A CN 201910842179 A CN201910842179 A CN 201910842179A CN 110467665 A CN110467665 A CN 110467665A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
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- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
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- G—PHYSICS
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- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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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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- G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
- G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/28—Neurological disorders
- G01N2800/2871—Cerebrovascular disorders, e.g. stroke, cerebral infarct, cerebral haemorrhage, transient ischemic event
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Abstract
This application discloses a kind of specific antigen epitope peptide of Occludin 31kDa degradation fragment and its applications.The specific antigen epitope peptide includes amino acid sequence shown in 323-334 of SEQ ID No.1, antibody caused by its antigen being prepared is only capable of identification Occludin 31kDa degradation fragment, it cannot identify Occludin 55kDa degradation fragment and undegraded Occludin, good with specificity, detection limits low advantage.The present invention is extreme early, quickly, efficiently measurement acute ischemic cerebral apoplexy patients serum 31-kDa Occludin Fragment Levels, prediction acute ischemic cerebral apoplexy lead to treatment cerebral hemorrhage again, provide corresponding new detecting technique.
Description
Technical field
The present invention relates to field of biotechnology, specifically a kind of specific antigen of Occludin 31kDa degradation fragment
Epitope peptide and its application.
Background technique
At present clinically, no matter CT (computed Tomography, CT scan) or MRI
(Magnetic Resonance Imaging, magnetic resonance imaging), all can not be real for the degree of injury of blood-brain barrier before thrombolysis
Now accurate and quickly measurement.Once several serologic marker objects (including MMP-9 and cFn) of great expectations were sent to, are not only
The structural proteins of blood-brain barrier, and its serum levels is also influenced vulnerable to other factors, all in all, it is difficult to accurately reflect blood brain screen
Hinder degree of injury.
Occludin (i.e. Occludin albumen) is one of the key molecule to form blood-brain barrier " strip of paper used for sealing " effect.Largely grind
Study carefully the generation for showing Occludin degradation fragment, indicates that the impaired degree of blood-brain barrier is serious.We have discovered that ischemic 2
Hour can cause the Occludin fast degradation of brain microvessel endothelial cells in vitro；It is sent out in animal model and clinical test
It is existing: 1) in 4.5 hours after cerebral ischemia generation, 55kDa can successively to be detected in peripheral blood and two kinds of 31kDa different size of are closed
Lock protein degradation segment.2) 2 hours after ischemic, the raising of 55kDa Fragment Levels can be found in peripheral blood, but its serum is normal
Baseline level (i.e. before cerebral ischemia) is also high, and the rate of climb is slow.3) 4 hours after ischemic, 31kDa piece can be found in peripheral blood
Duan Shuiping is sharply increased suddenly, and serum normal baseline levels (i.e. before cerebral ischemia) are lower, and the threshold value that " all or none " sample is presented becomes
Change effect (document: Rong Pan et al.Blood Occludin Level as a Potential Biomarker for
Early Blood Brain Barrier Damage Following Ischemic Stroke.Sci Rep.2017；7:
40331).Therefore, Occludin 31kDa degradation fragment can be used as the biological marker of Blood Brain Barrier (BBB) permeability degree before evaluation thrombolysis
Object.Extreme early, quickly, efficiently measurement acute ischemic cerebral apoplexy patients serum Occludin 31kDa degradation fragment is horizontal, can be right
Predict that acute ischemic cerebral apoplexy leads to treatment cerebral hemorrhage again and provides reference.
Summary of the invention
The purpose of the present invention is to provide a kind of specific antigen epitope peptide of Occludin 31kDa degradation fragment and its answer
With antibody caused by the antigen be prepared as the specific antigen epitope peptide is only capable of identification Occludin 31kDa degradation piece
Section, cannot identify Occludin 55kDa degradation fragment and undegraded Occludin, have specificity good, and detection limits low advantage.
On the one hand, the present invention provides a kind of specific antigen epitope peptide of Occludin 31kDa degradation fragment, it includes
Amino acid sequence shown in 323-334 of SEQ ID No.1；
The amino acid sequence of the Occludin as shown in SEQ ID No.1, the Occludin 31kDa degradation fragment
Amino acid sequence is as shown in 321-522 of SEQ ID No.1.
On the other hand, the present invention provides a kind of specific antigens of Occludin 31kDa degradation fragment, by claim
Specific antigen epitope peptide described in 1 is prepared with carrier protein couplet.
It on the other hand, is by described the present invention provides a kind of specific antibody of Occludin 31kDa degradation fragment
The monoclonal antibody or polyclonal antibody that specific antigen is prepared.
On the other hand, the present invention provides the specific antigen epitope peptides, the specific antigen or described
Specific antibody is in preparing the product (reagent, kit, test strips etc.) for detecting Occludin 31kDa degradation fragment
Using；
Preferably, the sample of the detection is peripheral blood serum, it is furthermore preferred that the derived from peripheral blood is in acute ischemic
Patients with cerebral apoplexy.
In a kind of specific embodiment, the present invention provides a kind of glue for detecting Occludin 31kDa degradation fragment
Body gold immune chromatography test paper, the test paper include: the specific antibody, detection line and the nature controlling line of colloid gold label；
Further, in the specific antibody of the colloid gold label, the standard gold amount of the specific antibody
For 15-25 μ g/ml, preferably 20 μ g/ml, the partial size of the colloidal gold is 18-22nm；
Further, the detection line includes the specific antibody, it is preferred that the concentration of the antibody is 0.4-
0.8mg/mL, more preferable 0.6mg/mL；
Further, the nature controlling line includes immunoglobulin, it is preferred that the concentration of the immunoglobulin is 0.5-
1mg/mL, more preferable 0.8mg/mL.
On the other hand, the present invention provides the specific antigen epitope peptide, the specific antigen, the spies
The application of heterogenetic antibody, the immune chromatography test paper in detection Occludin 31kDa degradation fragment, it is preferred that the inspection
The sample of survey is peripheral blood serum, it is furthermore preferred that the derived from peripheral blood is in patients with acute ischemic cerebral stroke.
On the other hand, the present invention provides the specific antigen epitope peptide, the specific antigen, the spies
Heterogenetic antibody, the immune chromatography test paper are preparing product (reagent, examination for Blood Brain Barrier (BBB) permeability degree before detecting thrombolysis
Agent box, test strips etc.) in application.
The present invention has the beneficial effect that:
Cerebral hemorrhage is that the severe complication that cerebral apoplexy leads to treatment again at present clinically both can not accurate evaluation before the treatment
Risk of intracerebral hemorrhage also lacks and mitigates the effective means that cerebral hemorrhage has serious consequences.Occludin 31kDa drop provided by the invention
The specific antigen epitope peptide of segment is solved, antibody caused by the antigen being prepared is only capable of identification Occludin 31kDa drop
Segment is solved, cannot identify Occludin 55kDa degradation fragment and undegraded Occludin, has specificity good, detection limit is low
Advantage.Therefore, the present invention realizes life of the Occludin 31kDa degradation fragment as Blood Brain Barrier (BBB) permeability degree before evaluation thrombolysis
The purposes of object marker, and for extreme early, quickly, efficiently measurement acute ischemic cerebral apoplexy patients serum 31-kDa Occludin
Fragment Levels, prediction acute ischemic cerebral apoplexy lead to treatment cerebral hemorrhage again, provide corresponding new detecting technique.The present invention is early
Phase predicts that acute ischemic cerebral apoplexy leads to the risk for the treatment of cerebral hemorrhage again, improves the safety of acute ischemic cerebral apoplexy treatment,
Provide new strategy and means.
Detailed description of the invention
Fig. 1 is the sensitivity technique result of 4 colloidal gold immune chromatography test of embodiment.
Specific embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
Embodiment 1, the epitope peptide design of 31kDa Occludin degradation fragment and synthesis
Analyze people Occludin albumen (Occludin) amino acid sequence (shown in SEQ ID No.1), Occludin albumen
31kDa degradation fragment amino acid sequence (321-522 shown in SEQ ID No.1) and Occludin albumen 55kDa degradation
Three sections of amino acid sequences are chosen in fragment amino acid sequence position in Occludin protein 31 kDa degradation fragment amino acid sequence
As the candidate specific antigen epitope of Occludin protein 31 kDa degradation fragment, it is respectively synthesized to obtain three polypeptides, i.e. antigen
Epitope peptide T1, T2, T3, sequence difference are as follows:
323-334 shown in epitope peptide T1:SEQ ID No.1；
346-355 shown in epitope peptide T2:SEQ ID No.1；
363-373 shown in epitope peptide T3:SEQ ID No.1.
The preparation of embodiment 2, antigen and antibody
1 three obtained polypeptide (epitope peptide) of embodiment is connect with carrier protein KLH (hemocyanin) respectively, is made
It is standby to obtain three kinds of antigens with immunogenicity.
Using three kinds of antigens as immunizing antigen, using hypodermic injection immunizing rabbit, every 2 weeks booster immunizations one
It is secondary, it is immunized 4 times altogether, arteria carotis takes blood, obtains antiserum, then carry out purifying and obtain three kinds of antibody.
It is detection antigen with corresponding epitope peptide-BSA, to the antiserum containing corresponding antibodies point using ELISA method
Not carry out bioactivity, as a result up to 1:3000 or more.
Embodiment 3, Western blot detect antibody specificity
Take each portion of the blood serum sample of 3 patients with acute ischemic cerebral stroke peripheral bloods, carry out protein electrophorese separation and
Transferring film, then each sample is detected respectively with three kinds of antibody that embodiment 2 is prepared, it is anti-with Occludin Protein Detection
Body is control, and the results are shown in Table 1.
Table 1, Western blot detect antibody specificity result
Note: the "-" in table 1 indicates that result is feminine gender, and "+" indicates that result is the positive.
Table 1 is the results show that antibody KT-1 has identification Occludin 31kDa degradation fragment and cannot identify Occludin
The specificity of 55kDa degradation fragment and undegraded Occludin, antibody KT-2 and antibody KT-3 do not have identification Occludin
The specificity of 31kDa degradation fragment.
Embodiment 4, preparation quickly detect the colloidal gold immune chromatography test of Occludin 31kDa degradation fragment
1, the preparation of colloidal gold
The colloidal gold solution for using trisodium citrate reduction method to prepare partial size as 20nm.
2, the preparation of the specific antibody of colloid gold label
The colloidal gold solution that step 1 obtains is taken, 0.2M K is added2CO3Make the PH 8.2 of colloid gold particle；Add 200 μ
G antibody KT-1 stirs 10-20min；10%BSA1.2ml is added, 10-20min is stirred；Add 0.05ml 20%
PEG2000 stirs 8min；Then 4 DEG C it is 0.5 hour static, the specific antibody of colloid gold label is centrifuged, supernatant is abandoned, is used
0.2M PB pH7.4(0.2M NaH2PO4, 0.2M Na2HPO40.5g BSA is added after mixing in equivalent) it is settled to 2ml, obtain glue
The specific antibody of body gold label, the standard gold amount of the specific antibody are 20 μ g/ml.
3, the assembling of colloidal gold immune chromatography test
The specific antibody solution of colloid gold label prepared by step 2 is sprayed on glass fibre membrane, 37 DEG C of constant temperature dryings.
0.6mg/mL antibody KT-1 and 0.8mg/mL goat anti-rabbit igg is fixed on NC film, respectively as detection line (T line) and nature controlling line
(C line), line-to-line are placed in 37 DEG C of vacuum constant temperature drying boxes dry 2h or more every 0.8cm.By sample pad (2cm long), colloidal gold
Bonding pad (5mm long), NC film and water absorption pad (2cm long) are successively affixed in bottom plate PVC board along its length, then are cut into width
0.5cm/ item obtains the colloidal gold immune chromatography test of quickly detection Occludin 31kDa degradation fragment.
4, the detection method (double antibody sandwich method) and criterion of colloidal gold immune chromatography test
The test paper made in step 3 is taken, sample pad one end is immersed in the container for filling test serum, observation in 15min
As a result.If occurring two red bands at test strips nature controlling line and detection line, show that sample for the positive, contains Occludin
31kDa degradation fragment；If only having in test strips and occurring a red stripes at nature controlling line, show sample for feminine gender, without containing locking
Protein 31 kDa degradation fragment；Illustrate that test strips fail if not occurring red stripes at the nature controlling line.
5, test strips sensitivity experiments
The test strips prepared with step 3 detect the standard solution configured with 0.01M PBS buffer solution (respectively containing real simultaneously
0,0.1,1,10, the 100 and 1000ng/mL of antigen being prepared in example 2 by epitope peptide T1 is applied (with Occludin 31kDa
Degradation fragment meter)), there is susceptibility of the minimum concentration as test strips of positive signal.
As a result as shown in Figure 1, showing the standard solution of test strips detection various concentration prepared by step 3, work as antigen concentration
Detection line is still as it can be seen that show that the test strip sensitivity is good when for 1ng/mL.
It is of different shades according to standard solution in T detection line position, the standard solution of settable various concentration gradient,
To estimate the Occludin 31kDa degradation fragment concentration range in sample to be tested.
Simultaneously according to method same as described above, colloidal gold immunochromatographimethod is prepared using antibody KT-2 and antibody KT-3 respectively
Test paper.
The application of embodiment 5, colloidal gold immune chromatography test before detecting thrombolysis in Blood Brain Barrier (BBB) permeability degree takes thrombolysis
The peripheral blood serum of preceding acute ischemia patients with cerebral apoplexy and Healthy People, while blood-brain barrier's damage is obtained by conventional method
Degree is detected.
Using the colloidal gold immune chromatography test of embodiment 4 prepared using antibody KT-1, blood serum sample is detected,
Will test result and standard solution (respectively containing the antigen 0 being prepared in embodiment 2 by epitope peptide T1,1,2,3,4,
5,6,7,8,9,10ng/mL (in terms of Occludin 31kDa degradation fragment)) testing result be compared, as a result such as 2 institute of table
Show.
Occludin 31kDa degradation fragment concentration in table 2, sample to be tested
| Sample | Patient 1 | Patient 2 | Patient 3 | Patient 4 | Healthy People |
| Concentration ng/mL | 9 | 8 | 10 | 8 | 1 |
By with patient's Occludin 31kDa degradation fragment actual concentrations (protein electrophoresis separates, Western is detected and
Determination of protein concentration), practical Blood Brain Barrier (BBB) permeability degree situation compares, the results showed that, antibody KT-1 system is used shown in table 2
The actual concentrations of Occludin 31kDa degradation fragment concentration and Occludin 31kDa degradation fragment that standby test paper detects
It is consistent, and consistent with Blood Brain Barrier (BBB) permeability degree, i.e., concentration is higher, and Blood Brain Barrier (BBB) permeability degree is more serious.
Above-mentioned identical blood serum sample is carried out using the test paper of embodiment 4 prepared by antibody KT-2 and antibody KT-3 simultaneously
Detection, since it shows that result is Occludin 31kDa degradation fragment, Occludin 55kDa degradation fragment and undegradable closes
Albumen is locked, testing result is apparently higher than the actual concentrations of patient's Occludin 31kDa degradation fragment, and testing result is in Healthy People
Concentration difference between patient is unobvious, if being compared with the testing result of aforesaid standards solution or color, it may appear that false
It is positive.
The content being not described in detail in this specification belongs to the prior art well known to professional and technical personnel in the field.More than
Described is only embodiments herein, is not intended to limit this application.To those skilled in the art, the application can
To there is various modifications and variations.All any modification, equivalent replacement, improvement and so within the spirit and principles of the present application,
It should be included within the scope of the claims of this application.
Sequence table
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Met Ser Ser Arg Pro Leu Glu Ser Pro Pro Pro Tyr Arg Pro Asp Glu
1 5 10 15
Phe Lys Pro Asn His Tyr Ala Pro Ser Asn Asp Ile Tyr Gly Gly Glu
20 25 30
Met His Val Arg Pro Met Leu Ser Gln Pro Ala Tyr Ser Phe Tyr Pro
35 40 45
Glu Asp Glu Ile Leu His Phe Tyr Lys Trp Thr Ser Pro Pro Gly Val
50 55 60
Ile Arg Ile Leu Ser Met Leu Ile Ile Val Met Cys Ile Ala Ile Phe
65 70 75 80
Ala Cys Val Ala Ser Thr Leu Ala Trp Asp Arg Gly Tyr Gly Thr Ser
85 90 95
Leu Leu Gly Gly Ser Val Gly Tyr Pro Tyr Gly Gly Ser Gly Phe Gly
100 105 110
Ser Tyr Gly Ser Gly Tyr Gly Tyr Gly Tyr Gly Tyr Gly Tyr Gly Tyr
115 120 125
Gly Gly Tyr Thr Asp Pro Arg Ala Ala Lys Gly Phe Met Leu Ala Met
130 135 140
Ala Ala Phe Cys Phe Ile Ala Ala Leu Val Ile Phe Val Thr Ser Val
145 150 155 160
Ile Arg Ser Glu Met Ser Arg Thr Arg Arg Tyr Tyr Leu Ser Val Ile
165 170 175
Ile Val Ser Ala Ile Leu Gly Ile Met Val Phe Ile Ala Thr Ile Val
180 185 190
Tyr Ile Met Gly Val Asn Pro Thr Ala Gln Ser Ser Gly Ser Leu Tyr
195 200 205
Gly Ser Gln Ile Tyr Ala Leu Cys Asn Gln Phe Tyr Thr Pro Ala Ala
210 215 220
Thr Gly Leu Tyr Val Asp Gln Tyr Leu Tyr His Tyr Cys Val Val Asp
225 230 235 240
Pro Gln Glu Ala Ile Ala Ile Val Leu Gly Phe Met Ile Ile Val Ala
245 250 255
Phe Ala Leu Ile Ile Phe Phe Ala Val Lys Thr Arg Arg Lys Met Asp
260 265 270
Arg Tyr Asp Lys Ser Asn Ile Leu Trp Asp Lys Glu His Ile Tyr Asp
275 280 285
Glu Gln Pro Pro Asn Val Glu Glu Trp Val Lys Asn Val Ser Ala Gly
290 295 300
Thr Gln Asp Val Pro Ser Pro Pro Ser Asp Tyr Val Glu Arg Val Asp
305 310 315 320
Ser Pro Met Ala Tyr Ser Ser Asn Gly Lys Val Asn Asp Lys Arg Phe
325 330 335
Tyr Pro Glu Ser Ser Tyr Lys Ser Thr Pro Val Pro Glu Val Val Gln
340 345 350
Glu Leu Pro Leu Thr Ser Pro Val Asp Asp Phe Arg Gln Pro Arg Tyr
355 360 365
Ser Ser Gly Gly Asn Phe Glu Thr Pro Ser Lys Arg Ala Pro Ala Lys
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Gly Arg Ala Gly Arg Ser Lys Arg Thr Glu Gln Asp His Tyr Glu Thr
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Asp Tyr Thr Thr Gly Gly Glu Ser Cys Asp Glu Leu Glu Glu Asp Trp
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Lys Arg Asn Phe Asp Thr Gly Leu Gln Glu Tyr Lys Ser Leu Gln Ser
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Glu Leu Asp Glu Ile Asn Lys Glu Leu Ser Arg Leu Asp Lys Glu Leu
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Asp Asp Tyr Arg Glu Glu Ser Glu Glu Tyr Met Ala Ala Ala Asp Glu
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Tyr Asn Arg Leu Lys Gln Val Lys Gly Ser Ala Asp Tyr Lys Ser Lys
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Lys Asn His Cys Lys Gln Leu Lys Ser Lys Leu Ser His Ile Lys Lys
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Met Val Gly Asp Tyr Asp Arg Gln Lys Thr
515 520
Claims (10)
1. the specific antigen epitope peptide of Occludin 31kDa degradation fragment, it is characterised in that: comprising SEQ ID No.1
Amino acid sequence shown in 323-334；
The amino acid sequence of the Occludin is as shown in SEQ ID No.1, the amino of the Occludin 31kDa degradation fragment
Acid sequence is as shown in 321-522 of SEQ ID No.1.
2. the specific antigen of Occludin 31kDa degradation fragment, it is characterised in that: anti-by specificity described in claim 1
Former epitope peptide is prepared with carrier protein couplet.
3. the specific antibody of Occludin 31kDa degradation fragment, it is characterised in that: be by specificity as claimed in claim 2
The monoclonal antibody or polyclonal antibody that antigen is prepared.
4. specific antigen epitope peptide described in claim 1, specific antigen as claimed in claim 2 or claim 3 institute
The specific antibody stated is preparing the application in the product for detecting Occludin 31kDa degradation fragment；
Preferably, the sample of the detection is peripheral blood serum, it is furthermore preferred that the derived from peripheral blood is in acute ischemic brain soldier
Middle patient.
5. a kind of colloidal gold immune chromatography test for detecting Occludin 31kDa degradation fragment, it is characterised in that: the test paper packet
It includes: specific antibody as claimed in claim 3, detection line and the nature controlling line of colloid gold label.
6. colloidal gold immune chromatography test according to claim 5, it is characterised in that: the right of the colloid gold label is wanted
In specific antibody described in asking 3, the standard gold amount of the specific antibody is 15-25 μ g/ml, preferably 20 μ g/ml, the glue
The partial size of body gold is 18-22nm.
7. colloidal gold immune chromatography test according to claim 5 or 6, it is characterised in that: the detection line includes right
It is required that specific antibody described in 3, it is preferred that the concentration of the antibody is 0.4-0.8mg/mL, more preferable 0.6mg/mL.
8. according to the colloidal gold immune chromatography test any in claim 5-7, it is characterised in that: the nature controlling line includes
Immunoglobulin, it is preferred that the concentration of the immunoglobulin is 0.5-1mg/mL, more preferable 0.8mg/mL.
9. described in specific antigen epitope peptide described in claim 1, specific antigen as claimed in claim 2, claim 3
Specific antibody, in claim 5-8 any immune chromatography test paper in detection Occludin 31kDa degradation fragment
Application, it is preferred that the sample of the detection be peripheral blood serum, it is furthermore preferred that the derived from peripheral blood is in acute ischemic
Patients with cerebral apoplexy.
10. specific antigen epitope peptide described in claim 1, specific antigen as claimed in claim 2, claim 3 institute
Any immune chromatography test paper is being prepared for detecting blood brain screen before thrombolysis in the specific antibody stated, claim 5-8
Hinder the application in the product of degree of injury.
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| CN112147323A (en) * | 2020-09-07 | 2020-12-29 | 深圳市第二人民医院 | Test strip for detecting 31-kDa Occludin after thrombolysis and preparation method and application thereof |
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| RONG PAN等: "Blood Occludin Level as a Potential Biomarker for Early Blood Brain Barrier Damage Following Ischemic Stroke", 《NATURE》 * |
| 杨可等: "闭合蛋白在脑缺血中的作用", 《国际脑血管病杂志》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112147323A (en) * | 2020-09-07 | 2020-12-29 | 深圳市第二人民医院 | Test strip for detecting 31-kDa Occludin after thrombolysis and preparation method and application thereof |
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| CN110467665B (en) | 2021-05-04 |
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