CN107356758B - A kind of lung cancer detection kit and its application method - Google Patents
A kind of lung cancer detection kit and its application method Download PDFInfo
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- CN107356758B CN107356758B CN201710743671.0A CN201710743671A CN107356758B CN 107356758 B CN107356758 B CN 107356758B CN 201710743671 A CN201710743671 A CN 201710743671A CN 107356758 B CN107356758 B CN 107356758B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- 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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57423—Specifically defined cancers of lung
Abstract
The present invention relates to biologic medical fields, in particular to a kind of lung cancer detection kit and its application method.The kit includes: solid phase carrier and the second solution of magnetic nanoparticles, and the solid phase carrier fixation is coated with the first magnetic nanoparticle；First magnetic nanoparticle is coated with GM1 capturing agent, and second magnetic nanoparticle is coated with GM1 detection agent；Wherein, the GM1 capturing agent and GM1 detection agent are the pairing antibody in conjunction with GM1；Or the GM1 capturing agent is GM1 antibody, the GM1 detection agent is amino acid sequence GM-TAG albumen as shown in SEQ ID NO:1；Or the GM1 capturing agent is the GM-TAG albumen, the GM1 detection agent is GM1 antibody.Detection instrument provided by the present invention is suitble to extensive manufacture, easy to use, is suitble to widely available；There is the ability of early detection lung cancer, and indagation is highly reliable.
Description
Technical field
The present invention relates to biologic medical fields, in particular to a kind of lung cancer detection kit and its application method.
Background technique
Lung cancer (Lung cancer, LC) is most important cancer in the world, and about 1,300,000 people die of lung cancer every year.Entirely
The Chen Wanqing of tumour Register, state is equal to has delivered 2015 China on January 25th, 2016 on CA Cancer J Clin magazine
Cancer statistical data.It was reported that China has 429.2 ten thousand cancer new cases, 281.4 ten thousand cancer mortalities for 2015.Lung cancer
As the most common cancer and the first cause of cancer mortality.
Due to a variety of environmental changes, including aging, health threat relevant to lung cancer and death may also be will increase.Root
According to cytology and origin of cell, lung cancer can be divided into multiple subspecies classes.Main Types include non-small cell carcinoma (NSCLC) and
Small cell carcinoma (SCLC).
Although radiology, surgery and chemotherapy obtain progress, 5 years survival rates of Past 30 Years lung cancer do not have substantially
It changes, only 15% patient 5-year Survival or 5 years or more after initial diagnosis.These low survival rates mainly by
The diagnosis of early stage is hampered in some simultaneous illnesss, limits the effect of therapy intervention.
Current small throughput diagnostic method includes sputum smear, chest x-ray and CT scan.These methods are only when patient has
Characteristic symptom, such as long-term cough, spit, hemoptysis, uncomfortable, pain or loss of weight, and hospital is gone to just to use when seeing the doctor,
Thus these methods are not appropriate for carrying out high flux examination to a large amount of crowds.In addition, these methods false positive with higher, it must
Subsequent invasive means, such as bronchoscope or tissue biopsy must be cooperated, made a definite diagnosis.Cytology detection is based on cell shape
State, structure and anatomy, can be used hematoxylin and eosin dyeing is handled.The means are widely used in medical diagnosis,
And by the gold standard as screening for cancer biopsy.Then hematoxylin uses the solution counterstain of eosin Y, daybreak to nuclear targeting
Red Y is dyed to cytoplasm fraction, generally includes intracellular and extracellular protein.The colouring method can be used for critical tumorogenic egg
White in-site detecting；Such as epithelial growth factor receptor (EGFR), Cyclin D1 or K-Kas.Anticancer egg is directed to these
The relevant ligase of white antibody, such as horseradish peroxidase (HRP) or alkaline phosphatase (AP) or fluorescent dye such as fluorescein
It can be used for finding the amount or cell position of any oncogene in biopsy.Similarly, using tumor inhibitor antibody, such as p53
Or Retinoblastoma Protein (pRB), it can be used for finding the cell position of weak sign or change；Such methods can mention
A possibility that showing canceration is increasing.Although such methods comparative maturity, speed are slower, it is impossible to be used in extensive high-throughput sieve
It looks into.
Many methods wish to more effectively diagnosing.For example, for the nucleic acid that can encode albumen, using specific
Primer or probe carry out diagnosing, and numerical value is believed to state (i.e. phosphatidylinositols-glycan specificity of instruction lung cancer
Phospholipase D；GPLD1).However, it is this use albumen as the method for pulmonary cancer diagnosis biomarker there are no commercialized elder generations
Example.It is any to be difficult to detect early stage using the method that blood is detected since the early stage of lung cancer is scattered in the different location of lung
Lung cancer because blood usually reflect be the entire circulatory system situation.Therefore, it is necessary to research and develop a kind of inexpensive and effective side
Method is used for quick diagnosis, and is easy to be commonly used.
In view of this, the present invention is specifically proposed.
Summary of the invention
The purpose of the present invention is to provide a kind of lung cancer detection kit and its application methods.
In order to realize above-mentioned purpose of the invention, the following technical scheme is adopted:
The present invention relates to a kind of lung cancer detection kit, the kit includes:
Solid phase carrier and the second solution of magnetic nanoparticles, the solid phase carrier fixation are coated with the first magnetic Nano
Grain；
First magnetic nanoparticle is coated with GM1 capturing agent, and second magnetic nanoparticle is coated with GM1 detection
Agent；
Wherein, the GM1 capturing agent and GM1 detection agent are the pairing antibody in conjunction with GM1；
Or the GM1 capturing agent is GM1 antibody, the GM1 detection agent is amino acid sequence as shown in SEQ ID NO:1
GM-TAG albumen；
Or the GM1 capturing agent is the GM-TAG albumen, the GM1 detection agent is GM1 antibody.
Wherein GM-TAG albumen and antibody are incorporated into the different epitopes of the GM1.
Kit provided by the invention is for quickly and easily diagnosing.The using effect of the kit have it is easy,
Quickly, the advantages that sensitive, special, stable.
Based on the scientific research before applicant, applicant is using phlegm (from bottom respiratory tract, especially trachea and bronchus
The mucus of expectoration) it is diagnosed.This becomes detection simply and readily.Sino-British international research before applicant is true
It accepts some chemical markers objects, this examines method provided by the invention faster economically relative to other methods
Disconnected lung cancer, while patient there will not be sense of discomfort.Based on these biomarkers, applicant develops diagnostic tool.
In the research before applicant, mass spectrograph platform can easily establish database from sputum sample, to these
The analysis of data, it can be found that clinically relevant data.It, can be by health using principal component analysis and high intension analysis method
Control group is distinguished from those patients for needing to carry out further lung cancer inspection.Although then these not all diseases
People is confirmed to be patients with lung cancer, and there are those doubtful patients with lung cancer those to think symptom relevant to cancer by doctor, it should
It is divided into " non-health ".In order to further confirm that the biomarker for distinguishing cancer and non-cancer, more complicated data are needed
Digging tool.ROC curve is the tool of a widely used judgement biomarker effect.ROC curve show true positives with
Relationship between false positive, so as to determine the using effect of a metabolin.Area under ROC curve can represent one
The classification performance of parameter: lung cancer and non-lung cancer.It is analyzed by using ROCCET, applicant have determined that a series of LTQ-MS- generation
Thank to object, the AUC value of these metabolins is greater than 0.8, this is one and confirms the whether useful critical value of the substance.These metabolins exist
Concentration is relatively high in cancer sputum sample.
GM1 (monosialoganglioside) be the gangliosides containing a sialic acid, also known as single sialic acid four
Sugar ganglioside, also known as sialic acid glycosphingolipid, molecular weight 1574.GM1 is mainly used for nerve fiber research, but can be
Find that they participate in cell-ECM identification, cell-matrix attachment, cell growth and cell differentiation in most cell types.Shen
It asks someone to have been found that the key organism marker that concentration increase of the glycosphingolipid in phlegm is detection of early lung cancer, accuracy rate exists
80% or more.Other have similar application, such as blend with hemocyanin and detect small cell carcinoma in tissue of patient biopsy.
One important feature of glycosphingolipid be it can with certain bacterial pathogens generate certain toxin in motif in conjunction with,
Such as the B subunit of cholera toxin.This is pathogenic associated with cholera, but GM1 binding site can be with B subunit protein
Rest part separation, and lose pathogenic effects.Similar intestines poison an important factor for there are also whole world morbidity and mortality
Disposition Escherichia coli (ETEC).The Major Virulence Factors of ETEC release are heat labile enterotoxin LT, structurally and functionally
Similar to cholera toxin.LTB combination glycosphingolipid, the host receptor of toxin, but it is mutual with A type blood glucose and e. coli lipopolysaccharide
Effect has also in the past decade been identified.
Applicant is it has been investigated that can with the Protein G M-TAG in conjunction with GM1 high-affinity, it should be noted that the protein
There is histidine tag in the end C- (shown in SEQ ID NO:1), it means that its separation is economical.
The recombinant protein of polyhistidine label can easily express in Escherichia coli.Bacterium can be obtained by centrifugation,
And gained cell precipitate is cracked to discharge cell protein.After being cultivated in the affine resin containing cobalt ions, it will mark
Separation of Proteins, polyhistidine tag is in connection with high-affinity.Resin is washed out to remove and not interact
Protein.Then GM-TAG can be further applied as described below.
Preferably, lung cancer detection kit as described above, first magnetic nanoparticle and the second magnetic Nano
Grain is Fe3O4Magnetic nanoparticle.
Magnetic nanoparticle row can be coated with the core-shell structure of boiomacromolecule, not only have at centered on magnetic material
Good magnetic conductance tropism, it may have good bio-intermiscibility can be immunoreacted in conjunction with protein.
Preferably, lung cancer detection kit as described above, first magnetic nanoparticle and the second magnetic Nano
The partial size of grain is 10~60nm；Preferably 15~40nm；More preferably 18~25nm.
The partial size of magnetic nano-particle has very big influence to the coupling quantity of capturing agent or detection agent, to affect
The sensitivity of end reaction.
Preferably, lung cancer detection kit as described above, the kit further include eluant, eluent；
Preferably, the eluant, eluent is PBS.
Preferably, lung cancer detection kit as described above, the kit further include diluents；
Preferably, the diluents are PBST.
PBST solution of the present invention is the PBS solution containing 5%Tween-20.
Preferably, lung cancer detection kit as described above, the kit further include magnetometer.
Preferably, lung cancer detection kit as described above, the solid phase carrier are EP pipe or tissue culture plate.
Preferably, lung cancer detection kit as described above, the kit further include positive control, the positive control
For the GM1 solution of known concentration.
According to an aspect of the present invention, it is detected the invention further relates to a kind of using lung cancer detection kit as described above
The method of GM1 albumen, comprising:
1) sample to be detected is added drop-wise in solid phase carrier and is incubated for by；
2) is eluted extra by sample to be detected, and the second solution of magnetic nanoparticles is added and is incubated for；
3) after elutes the second extra magnetic nanoparticle, by solid phase carrier, with magnetometer detection magnetic force signal, and will
Gained signal value substitutes into the magnetic force signal-GM1 concentration standard curve pre-established and GM1 in the sample to be detected is calculated
Content.
Preferably, method as described above, the sample to be detected are phlegm or saliva.
Compared with prior art, the invention has the benefit that
1), detection instrument provided by the present invention is suitble to extensive manufacture；
2), detection instrument provided by the present invention is easy to use, can be used as daily screening means, can popularize on a large scale
It uses；
3), detection instrument provided by the present invention has the ability of early detection lung cancer；
4), detection instrument provided by the present invention has the higher indagation reliability more than 80% or more.
Detailed description of the invention
It, below will be to specific in order to illustrate more clearly of the specific embodiment of the invention or technical solution in the prior art
Embodiment or attached drawing needed to be used in the description of the prior art be briefly described, it should be apparent that, it is described below
Attached drawing is some embodiments of the present invention, for those of ordinary skill in the art, before not making the creative labor
It puts, is also possible to obtain other drawings based on these drawings.
Fig. 1 is magnetic force signal-GM1 concentration standard curve schematic diagram；
Fig. 2 is the embodiment schematic diagram of embodiment 1；Wherein, GM1 is abbreviated as GM；
Fig. 3 is the embodiment schematic diagram of embodiment 2；Wherein, GM1 is abbreviated as GM.
Specific embodiment
The present invention provides a kind of tool based on GM-TAG, this tool uses nano particle detection as based on magnetic force
A part of the platform of meter.
The tool is based on magnetometer, such platform can have handset kind.When electric current passes through a small coil, magnetometer
Generate a magnetic field.If magnetic-particle is placed into this magnetic field, the influence to magnetic field is proportional to the quantity of magnetic-particle.This
Kind influences to perceive by highly sensitive magnetometer and for diagnosing.Magnetometer can provide knot in minutes
Fruit, and usually used ELISA method needs several hours to provide result.
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specific
Condition person carries out according to conventional conditions or manufacturer's recommended conditions.Reagents or instruments used without specified manufacturer is
The conventional products that can be obtained by commercially available purchase.
Embodiment 1
The application method of detection instrument provided by the present invention is as follows:
1) will be added drop-wise in solid phase carrier after Sample Dilution to be detected and be incubated for；
2) .PBS is eluted extra by sample to be detected, and the second solution of magnetic nanoparticles is added and is incubated for；
3) after elutes the second extra magnetic nanoparticle, the magnetic-particle on solid phase carrier is eluted, is examined with magnetometer
Survey magnetic force signal, and gained signal value substituted into the magnetic force signal-GM1 concentration standard curve that pre-establishes be calculated it is described
The content of GM1 in sample to be detected.
Magnetic force signal-GM1 concentration standard curve it is consistent when establishing mode with normal detection, detect the GM1 of various concentration
The magnetism intensity of standard items, and make standard curve (Fig. 1).
Magnetic nanoparticle is related to human body GM1 antibody.When there are GM1, this will be acted in capture antibody and with GM1
Detection antibody between formed a sandwich region.Curent change in coil to and the relevant magnetic nanoparticle number of antibody
Amount is related.GM1 is more in sample, just has more nano particles and magnetic fields (Fig. 2).When there is no GM1, can observe
To lower background interference.Turn off magnetic field, all antibody can all be removed in magnetometer, so as to measure next sample
This.Second magnetic coil can be installed, detection magnetic force signal is carried out, confirmation container is clean.
The concentration of GM1 can indicate the size of lung-cancer-risk.
Embodiment 2
In the present embodiment, a kind of novel kit improved for embodiment 1 is provided.It is fixed to be coated on admittedly
GM1 capture antibody on phase carrier replaces (Fig. 2) by GM-TAG albumen.The binding site and " free " of GM-TAG and GM1 molecule
GM1 detection antibody difference.Therefore, for the detection position of GM1, there is no competitions.This will enhance GM-TAG, GM1-GM1 inspection
Survey the sensibility in antibody sandwich region.Other aspects, the two tools are identical.
Embodiment 3
In the present embodiment, a kind of novel kit improved for embodiment 1 is provided.The GM1 of " free "
Detection antibody is replaced by GM-TAG albumen.The GM1 of binding site and GM1 the capture antibody of GM-TAG and GM1 molecule detects antibody
Difference.Therefore, for the catch position of GM1, there is no competitions.This will enhance GM-TAG, and GM1-GM1 captures antibody sandwich
The sensibility in region.Other aspects, the two tools are identical.
Experimental example
The technology is tested using the sample of Britain and China respectively at present.Wherein Britain's sample totally 67 people,
Including 33 normal healthy controls samples.It include lung cancer and non-lung cancer patient in remaining 34 people.Using our method, to lung
Cancer/non-lung cancer accuracy rate of diagnosis has integrally reached 80%.Chinese sample totally 50 people, including 15 normal healthy controls samples.
It using our method, is compared by the marker with confirmation, lung cancer/non-lung cancer accuracy rate of diagnosis is equally reached
80% level.It is worth noting that, in these samples, having reached 100% to the accuracy rate of diagnosis of small cell carcinoma.
1: one 45 years old male patient of case tells his deep coughs of doctor, shortness of breath and fever.Doctor has found him
Average 20 cigarettes of suction in one day, and him is required to provide sputum sample, it is detected using the method for the invention.Doctor can be to the disease
People makes a definite diagnosis.Using the technology, entire interval between diagnosis was less than 24 hours.
2: one 55 years old patients of case are due to pectoralgia, cough, spit and smoking history and be hospitalized.Doctor requires her to provide
Sputum sample, and detected using our method.This method by patient's sputum sample compound with we have found that marker
It contrasts.According to comparing result, which thinks patient with 2 phase lung cancer.Doctor then passes through other methods, including chest X
X ray fluoroscopy x etc. confirms the diagnostic result and provides corresponding treatment.
3: one 72 years old males of case, there is smoking history, due to the weight loss of unknown cause, lasting cough, and Yi Jishang
It is relatively difficult when stair, go to community hospital to check.He provide sputum samples for doctor's requirement.It is detected using our method,
By the comparison with marker, it is found that patient's lung cancer is positive.According to the testing result, which is sent to specially by Community Doctor
Hospital is further checked, it is thus identified that inspection result.
4: one 79 years old women of case, have smoking history, due to lasting cough, spit, and asthma, referring physician.Society
Qu doctor requires her to provide sputum sample.The sample is compared using our method with marker.Lung cancer is negative as the result is shown.
Community Doctor is sent to special hospital's division of respiratory disease in order to ensure not mistaken diagnosis, by the patient, it was demonstrated that and it is other respiratory problems, and
Corresponding treatment is carried out.
5: one 84 years old patients of case, are once diagnosed as lung cancer.Due to there is the symptom of recurrence, diagnosed.
But conventional diagnostic means to the injury that may cause of patient with these serious symptoms and thus bring disease incidence and dead
Rate is died, not can be carried out these diagnosis.In order to monitor progression of the disease, which is required to provide sputum sample.Use our side
Method, we quickly can provide accurate detection as a result, referring to for doctor, and formulate corresponding treatment method.
6: one 77 years old female patients of case have smoking history, the weight loss of unknown cause, asthma, and can not grow
Distance proceed on foot community hospital diagnosis.Community Doctor acquires sputum sample, and is detected using our method, finds the disease
Human lung cancer is positive, therefore submits special hospital to carry out immunotherapy targeted autoantibody the patient.
Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；To the greatest extent
Present invention has been described in detail with reference to the aforementioned embodiments for pipe, but those skilled in the art should understand that: its
It is still possible to modify the technical solutions described in the foregoing embodiments, or to some or all of the technical features
It is equivalently replaced；And these are modified or replaceed, various embodiments of the present invention skill that it does not separate the essence of the corresponding technical solution
The range of art scheme.
SEQUENCE LISTING
<110>Li Hongjun;The Anhui Han Ji;Land Yi Shiaer heavily fortified point De Muer (Mur, Luis Alejandro Jose)
<120>a kind of lung cancer detection kit and its application method
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 449
<212> PRT
<213>artificial sequence
<400> 1
Met Arg Gly Ser Met Ala Asn Ile Asn Asp Ser Lys Ile Leu Ser Leu
1 5 10 15
Gln Asn Arg Lys Asn Thr Leu Val Asp Thr Ser Gly Tyr Asn Ala Glu
20 25 30
Val Ser Glu Glu Gly Asp Val Gln Leu Asn Pro Ile Phe Pro Phe Asp
35 40 45
Phe Lys Leu Gly Ser Ser Gly Glu Asp Arg Gly Lys Val Ile Val Thr
50 55 60
Gln Asn Glu Asn Ile Val Tyr Asn Ser Met Tyr Glu Ser Phe Ser Ile
65 70 75 80
Ser Phe Trp Ile Arg Ile Asn Lys Trp Val Ser Asn Leu Pro Gly Tyr
85 90 95
Thr Ile Ile Asp Ser Val Lys Asn Asn Ser Gly Trp Ser Ile Gly Ile
100 105 110
Ile Ser Asn Phe Leu Val Phe Thr Leu Lys Gln Asn Glu Asp Ser Glu
115 120 125
Gln Ser Ile Asn Phe Ser Tyr Asp Ile Ser Asn Asn Ala Pro Gly Tyr
130 135 140
Asn Lys Trp Phe Phe Val Thr Val Thr Asn Asn Met Met Gly Asn Met
145 150 155 160
Lys Ile Tyr Ile Asn Gly Lys Leu Ile Asp Thr Ile Lys Val Lys Glu
165 170 175
Leu Thr Gly Ile Asn Phe Ser Lys Thr Ile Thr Phe Glu Ile Asn Lys
180 185 190
Ile Pro Asp Thr Gly Leu Ile Thr Ser Asp Ser Asp Asn Ile Asn Met
195 200 205
Trp Ile Arg Asp Phe Tyr Ile Phe Ala Lys Glu Leu Asp Gly Lys Asp
210 215 220
Ile Asn Ile Leu Phe Asn Ser Leu Gln Tyr Thr Asn Val Val Lys Asp
225 230 235 240
Tyr Trp Gly Asn Asp Leu Arg Tyr Asn Lys Glu Tyr Tyr Met Val Asn
245 250 255
Ile Asp Tyr Leu Asn Arg Tyr Met Tyr Ala Asn Ser Arg Gln Ile Val
260 265 270
Phe Asn Thr Arg Arg Asn Asn Asn Asp Phe Asn Glu Gly Tyr Lys Ile
275 280 285
Ile Ile Lys Arg Ile Arg Gly Asn Thr Asn Asp Thr Arg Val Arg Gly
290 295 300
Gly Asp Ile Leu Tyr Phe Asp Met Thr Ile Asn Asn Lys Ala Tyr Asn
305 310 315 320
Leu Phe Met Lys Asn Glu Thr Met Tyr Ala Asp Asn His Ser Thr Glu
325 330 335
Asp Ile Tyr Ala Ile Gly Leu Arg Glu Gln Thr Lys Asp Ile Asn Asp
340 345 350
Asn Ile Ile Phe Gln Ile Gln Pro Met Asn Asn Thr Tyr Tyr Tyr Ala
355 360 365
Ser Gln Ile Phe Lys Ser Asn Phe Asn Gly Glu Asn Ile Ser Gly Ile
370 375 380
Cys Ser Ile Gly Thr Tyr Arg Phe Arg Leu Gly Gly Asp Trp Tyr Arg
385 390 395 400
His Asn Tyr Leu Val Pro Thr Val Lys Gln Gly Asn Tyr Ala Ser Leu
405 410 415
Leu Glu Ser Thr Ser Thr His Trp Gly Phe Val Pro Val Ser Glu Pro
420 425 430
Gly Ser Ala Trp Ser His Pro Gln Phe Glu Lys His His His His His
435 440 445
His
Claims (12)
1. lung cancer detection kit is in the application for preparing pulmonary cancer diagnosis agent, which is characterized in that the kit includes:
Solid phase carrier and the second solution of magnetic nanoparticles, the solid phase carrier fixation are coated with the first magnetic nanoparticle；
First magnetic nanoparticle is coated with GM1 capturing agent, and second magnetic nanoparticle is coated with GM1 detection agent；
Wherein, the GM1 capturing agent is GM1 antibody, and the GM1 detection agent is amino acid sequence as shown in SEQ ID NO:1
GM-TAG albumen；
Or the GM1 capturing agent is the GM-TAG albumen, the GM1 detection agent is GM1 antibody.
2. application according to claim 1, which is characterized in that first magnetic nanoparticle and the second magnetic Nano
Grain is Fe3O4Magnetic nanoparticle.
3. application according to claim 1, which is characterized in that first magnetic nanoparticle and the second magnetic Nano
The partial size of grain is 10~60nm.
4. application according to claim 3, which is characterized in that first magnetic nanoparticle and the second magnetic Nano
The partial size of grain is 15~40nm.
5. application according to claim 4, which is characterized in that first magnetic nanoparticle and the second magnetic Nano
The partial size of grain is 18~25nm.
6. application according to claim 1, which is characterized in that the kit further includes eluant, eluent.
7. application according to claim 6, which is characterized in that the eluant, eluent is PBS.
8. application according to claim 1, which is characterized in that the kit further includes diluents.
9. application according to claim 8, which is characterized in that the diluents are PBST.
10. application according to claim 1, which is characterized in that the kit further includes magnetometer.
11. application according to claim 1, which is characterized in that the solid phase carrier is EP pipe or tissue culture plate.
12. application according to claim 1, which is characterized in that the kit further includes positive control, and the positive is right
According to the GM1 solution for known concentration.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1829916A (en) * | 2003-07-30 | 2006-09-06 | 皇家飞利浦电子股份有限公司 | Use of magnetic particles for determining binding between bioactive molecules |
| CN102597774A (en) * | 2009-09-23 | 2012-07-18 | 皇家飞利浦电子股份有限公司 | Binding assay with multiple magnetically labelled tracer binding agents |
| CN103930783A (en) * | 2011-11-16 | 2014-07-16 | 皇家飞利浦有限公司 | Particle repulsion to enhance surface contact in magnetic particle immunoassays |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63258493A (en) * | 1987-04-15 | 1988-10-25 | Mitsui Toatsu Chem Inc | Anti-ganglioside gm, monoclonal antibody, cell producing said antibody and reagent composed thereof |
| EP2541250A1 (en) * | 2011-06-30 | 2013-01-02 | Koninklijke Philips Electronics N.V. | Molecular architecture on magnetic particles for affinity assays with low non-specific binding |
-
2017
- 2017-08-25 CN CN201710743671.0A patent/CN107356758B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1829916A (en) * | 2003-07-30 | 2006-09-06 | 皇家飞利浦电子股份有限公司 | Use of magnetic particles for determining binding between bioactive molecules |
| CN102597774A (en) * | 2009-09-23 | 2012-07-18 | 皇家飞利浦电子股份有限公司 | Binding assay with multiple magnetically labelled tracer binding agents |
| CN103930783A (en) * | 2011-11-16 | 2014-07-16 | 皇家飞利浦有限公司 | Particle repulsion to enhance surface contact in magnetic particle immunoassays |
Non-Patent Citations (3)
| Title |
|---|
| Identification of Escherichia coli Heat-Labile Enterotoxin by Means of a Ganglioside Immunosorbent Assay(GM1-ELISA) Procedure;Ann-Mari Svennerholm et al;《CURRENT MICROBIOLOGY》;19781231;第1卷;第19–23页 * |
| Mutational analysis of the role of ADP-ribosylation activity and GM1-binding activity in the adjuvant properties of the Escherichia coli heat-labile enterotoxin towards intranasally administered keyhole limpet hemocyanin;Lolke de Haan et al;《Eur. J. Immunol.》;19981231;第28卷;第1243-1250页 * |
| The biological activity of botulinum neurotoxin type C is;Jasmin Strotmeier et al;《Molecular Microbiology》;20110602;第81卷(第1期);第143-156页 * |
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