CN107807241A - The magnetic microparticle chemiluminescence detection kit and preparation method of a kind of intravascular ErbB1 - Google Patents
The magnetic microparticle chemiluminescence detection kit and preparation method of a kind of intravascular ErbB1 Download PDFInfo
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- CN107807241A CN107807241A CN201711068672.6A CN201711068672A CN107807241A CN 107807241 A CN107807241 A CN 107807241A CN 201711068672 A CN201711068672 A CN 201711068672A CN 107807241 A CN107807241 A CN 107807241A
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Abstract
The invention discloses a kind of magnetic microparticle chemiluminescence detection kit of intravascular ErbB1 and preparation method.The kit includes：Coupling has intravascular ErbB1 detection antibody, intravascular ErbB1 calibration object, chemiluminescence preexciting liquid A, chemiluminescence exciting liquid B and the cleaning fluid of the magnetic particle of intravascular ErbB1 capture antibody, acridinium ester label.Kit of the present invention is using Magneto separate chemiluminescence as detection means, in combination with acridinium ester label technology.Direct chemoluminescence method high sensitivity that the present invention establishes, high specificity, accurate quick, detection time is short, and testing result has higher accuracy and repeatability, and the kit is applicable to various luminometer devices.
Description
Technical field
The invention belongs to technical field of immune assay, a kind of specifically intravascular ErbB1 is exempted from
Epidemic disease magnetic microparticle chemiluminescence detection kit and preparation method.
Background technology
Soluble vascular endothelial growth factor receptor1 (soluble vascular endothelial growth
Factor receptor-1, sVEGFR-1), also referred to as solvable fms samples EGFR-TK -1 (sFlt-1), sVEGFR-1 lacks
The 7th immunoglobulin like domain of VEGFR-1, lack intracellular tyrosine kinase domain, therefore it does not have cell signal to turn
Function is led, but it still has the ability combined with VEGF (VEGF), placenta growth factor (PIGF) high-affinity.
SVEGFR-l mainly plays the effect to anti-vegf biological function in vivo, and it is mainly manifested in：（1）Anti-angiogenesis, suppression
The VEGF-VEGFR-2 signals of Angiogensis processed；（2）The Vascular permeability of VEGF mediations is disturbed by VEGR-1 or VEGFR-2
Property；（3）Antiinflammatory action is produced by the activation and migration of the monocyte and macrophage that weaken VEGF-VEGFR-1 dependences.Greatly
Quantifier elimination also demonstrate that VEGF above-mentioned biological function, and such as in human body, sVEGFR-1 is proved to maintaining cornea without blood vessel
Aspect plays a significant role, in the molecule of numerous anti-angiogenesis for being present in cornea (angiostatin, Endostatin etc.),
SVEGFR-1 plays the effect for the Angiogensis for suppressing VEGFR.
Preeclampsia is a kind of peculiar and again common disease of gravid woman, and the incidence of disease is in the women of primigravid
7%~10%, on its pathogenesis at present compared with generally acknowledged theory may implantation shallow with placenta, placental perfusion reduce and nourish it is thin
Born of the same parents' hypoxic-ischemic is relevant, is gravid woman, puerpera and perinatal feruses morbidity and main causes of death.Its teiology mechanism is all the time
The study hotspot of obstetrics.Recent research finds that preeclampsia or even eclampsia are due to that trophocyte moves to uterine spiral
Arteries and veins infiltrated shallow, placenta blood vessel network and forms bad and extensive blood vessel endothelium injury, cause Placental ischemia, anoxic and trigger
, show that the sick generation is relevant with trophocyte, the growth of endothelial cell and function, sFlt-l is as vascular endothelial growth factor
Sub (VEGF) and PLGF direct mortifier, horizontal raise causes trophoblast and angiogenic growth in placenta in preeclampsia
It is bad, and then cause endothelial cell damage, and trigger a series of clinical symptoms, the early prediction of preeclampsia or be at present
Early detection, the clinical diagnosis and treatment aspect of early intervention still lack effective prevention and treatment means, for sFlt-1, PLGF and son
The correlative study of the clinical diagnosis of epilepsy early stage has received significant attention.
Up to the present, the method for soluble vascular endothelial growth factor receptor1 in serum mainly has：Enzyme linked immunological
Method（ELISA）.Enzyme linked immunosorbent assay (ELISA)（ELISA）Detection technique is to utilize specific immune response between Ag-Ab
Identify the target molecule in biofluid, and containing using the quantitative reaction sample to be tested target of the chromogenic reaction of enzyme-substrate
Amount.Although this method detection is cheap, quick, sensitivity is also inadequate, is only applicable to the detection and identification of micro substance.Cause
This, establishing a kind of method of detection sex hormone binding globulin effective, quick, simple, sensitive, anti-interference is high has very
Important meaning.
The present invention uses method as direct chemoluminescence method, using acridinium ester as chemiluminescent labels with obvious excellent
Gesture, it is mainly manifested in：Reaction does not need catalyst, as long as alkaline environment can be carried out, is swift in response, can complete catching reaction
Caused photon, background luminescence is low, and signal to noise ratio is high, and disturbing factor is few, and reagent stability is good, and system is simple, and exciting liquid cost is low,
Easily and protein bind, and photon yield is not reduced acridinium ester after being coupled.The Magnetism particulate immuno chemistry luminescence method that the present invention establishes is sensitive
Spend height, high specificity, accurate quick, detection time is short, testing result have higher accuracy with it is repeated.
The content of the invention
It is an object of the invention to provide the blood that a kind of sensitivity is higher, the reaction time is short, simple to operate, anti-interference is high
The magnetic microparticle chemiluminescence detection kit and preparation method of pipe endepidermis growth factor receptors 1.
To achieve the above object, the present invention provides following technical scheme：
The magnetic microparticle chemiluminescence detection kit and preparation method of a kind of intravascular ErbB1, institute of the present invention
The magnetic microparticle chemiluminescence method of offer detects the kit of intravascular ErbB1, and blood is coupled using magnetic particle
Pipe endepidermis growth factor receptors 1 captures antibody, the intravascular ErbB1 detection antibody of acridinium ester label.Reagent
Chemiluminescence preexciting liquid A, the chemistry that box also includes intravascular ErbB1 calibration object, above-mentioned acridinium ester acts on
Luminous exciting liquid B and cleaning fluid.
As the further scheme of the present invention, described magnetic particle directly can be caught with intravascular ErbB1
Antibody coupling is obtained, magnetic particle and Streptavidin can be also coupled, while use the intravascular EGF of biotin labeling
Acceptor 1 captures antibody.
As the further scheme of the present invention, the surface modification group of the magnetic particle is one or more active function groups
Group, including but not limited to carboxyl, amino, p-toluenesulfonyl or Streptavidin-biotin.
As the further scheme of the present invention, described chemiluminescent labels are acridinium ester, as NSP-DMAE-NHS,
NSP-SA-NHS etc..
As the further scheme of the present invention, described acridinium ester label is intravascular ErbB1 inspection
Survey antibody.
As the further scheme of the present invention, the intravascular ErbB1 detection of described acridinium ester label is anti-
The Na that the buffer solution of body is pH 8.0-11.0, concentration is 0.01-0.20 mol/L2CO3-NaHCO3Buffer solution.
As the further scheme of the present invention, described capture antibody is monoclonal antibody with detection antibody.
As the further scheme of the present invention, described calibration object be with the Tris containing 0.5-5.0% BSA or PBS or
HEPES buffer solution is matrix, adds intravascular ErbB1 sterling configuration and forms, calibration object form is liquid.
As the further scheme of the present invention, described intravascular ErbB1 calibration object solution concentration point
It is not：0 pg /mL、100.0 pg /mL、500.0 pg /mL、1000.0 pg /mL、2000.0 pg /mL、3500.0 pg
/mL。
As the further scheme of the present invention, described chemiluminescence preexciting liquid A is H2O2And HNO3Mixed liquor, its
Middle H2O2Mass fraction be 0.01-5.0%, HNO3Concentration be 0.01-1.0 mol/L.
As the further scheme of the present invention, described chemiluminescence exciting liquid B is the mixed of Triton X-100 and NaOH
Liquid is closed, wherein Triton X-100 mass fraction is 0.01-2.0%, and NaOH concentration is 0.05-1.0 mol/L.
As the further scheme of the present invention, described cleaning fluid is：PH 7.0-9.0, concentration are 5.0-50.0 mmol/
L Tris or HEPES or other solution, wherein containing the Tween-20 that mass fraction is 0.01-0.25%.
The principle of the present invention is to be combined the high degree of specificity of antibody-antigene reaction with the high sensitivity that acridinium ester lights
Get up, using photon caused by acridinium ester catching reaction to detect production concentration.
The advantage of the invention is that combining magnetic microparticle chemiluminescence technology using double antibody sandwich method, blood vessel endepidermis is determined
The content of growth factor receptors 1.Acridinium ester has a clear superiority as the direct chemiluminescence of label, is mainly manifested in：Reaction
Do not need catalyst, as long as alkaline environment can be carried out, be swift in response, can photon completely caused by catching reaction, background hair
Light is low, and signal to noise ratio is high, and disturbing factor is few, and reagent stability is good, and system is simple, and exciting liquid cost is low, and acridinium ester is easily and protein
It is coupled, and photon yield is not reduced after connection.
Embodiment
The present invention provides magnetic microparticle chemiluminescence detection kit and the preparation of a kind of intravascular ErbB1
Method, for make the purpose of the present invention, technical scheme and effect definitely, it is clear, the present invention is described in more detail below.
The present invention provides magnetic microparticle chemiluminescence detection kit and the preparation of a kind of intravascular ErbB1
Method, wherein, magnetic microparticle chemiluminescence method provided by the present invention detects the reagent of intravascular ErbB1
Box, intravascular ErbB1 is coupled using magnetic particle and captures antibody, the intravascular EGF of acridinium ester label
Acceptor 1 detects antibody.The chemistry that kit also includes intravascular ErbB1 calibration object, above-mentioned acridinium ester acts on
Luminous preexciting liquid A, chemiluminescence exciting liquid B and cleaning fluid.
Specifically, magnetic particle coupling suspension of the present invention captures antibody by intravascular ErbB1
Magnetic particle and the reagent buffer composition of coupling, wherein the concentration of intravascular ErbB1 capture antibody is 0.1-
2.0 mg/mL。
Specifically, the kernel of magnetic bead of the present invention is ferroso-ferric oxide, described magnetic particle can directly with intravascular table
Skin growth factor acceptor 1 captures antibody coupling, can also be coupled magnetic particle and Streptavidin, while use biotin labeling blood
Pipe endepidermis growth factor receptors 1 captures antibody.Magnetic bead can influence accuracy using the endless bulk deposition of preceding solid phase, therefore select
Shi Yingxuan good dispersions, the slow magnetic bead of sinking speed.
Specifically, for the present invention when preparing magnetic particle suspension, the coupled antibody buffer solution is pH 5.0-6.0, concentration
For 20-200 mmol/L MES buffer solutions.
Specifically, for the present invention when preparing magnetic particle suspension, the Block buffer is the buffer solution containing 1%BSA.
Specifically, the present invention prepare mark liquid-phase reagent when, the mark buffer solution is that pH 8.0-11.0, concentration are
0.01-0.20 mol/L Na2CO3-NaHCO3Buffer solution.
Specifically, calibration object of the present invention is to be with the Tris containing 0.5-5.0% BSA or PBS or HEPES buffer solution
Matrix, add intravascular ErbB1 sterling configuration and form, calibration object form is liquid.
Specifically, chemiluminescence preexciting liquid A of the present invention is H2O2And HNO3Mixed liquor, wherein H2O2Quality
Fraction is 0.01-5.0%, HNO3Concentration be 0.01-1.0 mol/L.
Specifically, chemiluminescence exciting liquid B of the present invention is Triton X-100 and NaOH mixed liquor, wherein
Triton X-100 mass fraction is 0.01-2.0%, and NaOH concentration is 0.05-1.0 mol/L.
Specifically, cleaning fluid of the present invention is：PH 7.0-9.0, concentration be 5.0-50.0 mmol/L Tris or
HEPES or other solution, wherein containing the Tween-20 that mass fraction is 0.01-0.25%.
Below by embodiment, the present invention is described in detail.
Embodiment 1：A kind of magnetic microparticle chemiluminescence kit of the described intravascular ErbB1 of detection
Establishment and preparation method, comprise the following steps：
1. the establishment of kit：
A kind of magnetic microparticle chemiluminescence kit for detecting intravascular ErbB1 is set up, it is contained following group
Point：
The intravascular ErbB1 monoclonal antibody of magnetic particle coupling；
The intravascular ErbB1 monoclonal antibody of another strain of acridinium ester label；
Intravascular ErbB1 serial standards solution；
Chemiluminescence preexciting liquid A and chemiluminescence exciting liquid B；
Cleaning fluid.
2. coupling has the preparation of the magnetic particle suspension of intravascular ErbB1 monoclonal antibody
（1）1 mg carboxyls magnetic particle is taken in 0.5 mL centrifuge tubes, adds the MES buffer solutions that 200 μ L concentration are 0.1 mol/L,
It is vortexed and mixes, be placed on magnetic frame, standing 5 min makes magnetic particle be separated with liquid, supernatant discarding, washs 3 times, adds 200
μ L MES（PH is 6.0）Buffer solution, it is vortexed.
（2）15 μ g intravascular ErbB1 monoclonal antibody is added, makes the mol ratio of carboxyl and antibody
For 150:1, it is vortexed, revolving reaction pipe, is incubated at room temperature 30 min.
（3）The coupling reagent EDC that 10 μ L concentration are 10 mg/mL is added, in being vortexed on rotatable reactor, is incubated at room temperature 2
h。
（4）Take 200 glycine buffers of the μ L containing 1%BSA（Concentration is 50 mmol/L）Closed, time 1h.
（5）Supernatant is removed, adds 200 μ L cleaning buffer solution（TBS+0.05%Tween-20）, wash 3 times.
（6）The above-mentioned magnetic particle suspension prepared is placed in 500 μ L and preserves liquid（It is 300 mmol/L glycine, 2% sweet
Oil, 5% sucrose）In, 2-8 DEG C of preservation.
3. the preparation of the intravascular ErbB1 monoclonal antibody liquid-phase reagent of acridinium ester label
（1）Purify intravascular ErbB1 monoclonal antibody：By the 100 intravascular ErbB1s of μ g
Monoclonal antibody is placed in bag filter, and bag filter is placed in the mark buffer solution not less than 1 L and dialysed, and during which buffer solution is more
Change 5 times, last time dialysed overnight, the Na that mark buffer solution is pH 9.8, concentration is 50 mmol/L2CO3-NaHCO3Buffering
Liquid.
（2）1.7 mg acridinium ester NSP-DMAE-NHS is weighed, is dissolved in 447 μ L anhydrous dimethyl formamides（DMF）In,
It is made into 6.5 mmol/L NSP-DMAE-NHS DMF solutions.
（3）500 μ L centrifugations will be placed in through the intravascular ErbB1 monoclonal antibody solution after dialysis
In pipe, 200 μ L mark buffer solutions are added, then add 759 μ L, 6.5 mmol/L NSP-DMAE-NHS DMF solutions（Keep away
Light reaction）The mol ratio for making acridinium ester and intravascular ErbB1 monoclonal antibody is 7.4:1, vibration mixes, room
Temperature 1 h of lower reaction, adds the lysine that 100 μ L concentration are 10 g/L, stands 15 min, make mark reaction terminating.
（4）Label NSP-DMAE-NHS-Ab and free NSP-DMAE-NHS passes through Sephadex G-50 posts（1×
25cm）Separation, the PBS for being 0.1mol/L with purification buffer pH 6.3, concentration are balanced and are eluted chromatographic column.
（5）Protein peak is detected with chromatograph in separation process, the chemiluminescence intensity and 280nm for measuring efflux respectively are inhaled
Shading value.
（6）The eluent that shading value is high and absorbance is big is collected, stored frozen is dispensed after adding 1% BSA.
4. the preparation of intravascular ErbB1 calibration object
Intravascular ErbB1 sterling is configured to sign concentration with the Tris-HCl buffer solutions containing 2% BSA
For 0 pg/mL, 100.0 pg/mL, 500.0 pg/mL, 1000.0 pg/mL, 2000.0 pg/mL, 3500.0 pg/
The calibration object Grad of mL totally 6 concentration.
5. chemiluminescence exciting liquid A, B preparation
（1）Chemiluminescence preexciting liquid A is by H2O2And HNO3Composition.Wherein, wherein H2O2Mass fraction be 1.5%, HNO3It is dense
Spend for 0.1 mol/L, be distributed into 20 mL/ branch with brown bottle, 2-8 DEG C saves backup.
（2）Chemiluminescence exciting liquid B is made up of Triton X-100 and NaOH mixed liquor.Wherein, Triton X-100
Mass fraction be 1.0%, NaOH concentration be 0.4 mol/L, be distributed into 20 mL/ branch with brown bottle, 2-8 DEG C saves backup.
6. the preparation of cleaning fluid
In the beaker for weighing 3.05g Tris, 8.775g NaCl to 1000 mL, it is 0.05% Tween- to add 1mL mass fractions
20, stir and evenly mix, constant volume, saved backup after pH is adjusted into 7.6.
Embodiment 2：Magnetic microparticle chemiluminescence detection kit using described intravascular ErbB1 is entered
Row detection the step of be：
（1）The μ L of sample to be tested 50 are added in reaction cup, add the magnetic particle coupling μ L of suspension 150, vibration mixes, 37 DEG C
It is incubated 8 min.
（2）Separation, clean 3 times, the reaction vessel after washing, which is fully vibrated, makes magnetic particle scatter.
（3）The μ L of acridinium ester label 150 are added into reaction cup, vibration mixes, 37 DEG C of 7 min of incubation.
（4）Separation, clean 3 times, the reaction vessel after washing, which is fully vibrated, makes magnetic particle scatter.
（5）100 μ L chemiluminescence exciting liquid B are added after adding 100 μ L chemiluminescence preexciting liquid A, 1 s, measure it
Relative luminous intensity, the content of the sample medium vessels endepidermis growth factor receptors 1 proportional pass of luminous intensity corresponding thereto
System.
Embodiment 3：The performance indications of kit
（1）Sensitivity for analysis
By the use of 0 pg/mL calibration objects in kit as sample to be tested, replication 20 times, the RLU values of 20 measurement results are drawn
（Relative light unit）, calculate its average value（M）And standard deviation（SD）, RLU values corresponding to M+2SD are drawn, according to zero-dose school
Concentration-RLU values result between quasi- product and adjacent calibration object carries out 2 regression fits and draws linear function, by M+2SD's
RLU values are brought into above-mentioned equation, obtain corresponding concentration value.By minimum detection limit detection method in detection scheme, it is repeated 3 times
Experiment, it is 89.4 pg/mL to the sensitivity for analysis of intravascular ErbB1 finally to obtain this kit.
（2）Precision
With intravascular ErbB1 kit test concentrations be (857.0 ± 163.5) pg/mL and (2600.0 ±
307.0) pg/mL sample, each concentration retest 10 times, kit precision is calculated, the results showed that kit CV is equal
Less than 5%.
（3）Stability
Intravascular ErbB1 detection kit is taken to carry out normal storage stability test, 2-8 DEG C of placement is pressed respectively
1,3,5,7,9,11,12,13,14,15 months time was detected；Uncap stability test respectively by 2-8 DEG C place 0 day, 7
My god, be measured within 14 days, 16 days, 18 days, 20 days, 22 days, 24 days, 26 days, 28 days, 30 days, 32 days.As a result intravascular table is shown
The detection kit of skin growth factor acceptor 1 is stored in 2-8 DEG C, in light protected environment, and the term of validity is 12 months.Uncap and be stored in 2-8
DEG C, in light protected environment, the term of validity is 30 days.
It is obvious to a person skilled in the art that the invention is not restricted to the details of above-mentioned one exemplary embodiment, Er Qie
In the case of without departing substantially from spirit or essential attributes of the invention, the present invention can be realized in other specific forms.Therefore, no matter
From the point of view of which point, embodiment all should be regarded as exemplary, and be nonrestrictive, the scope of the present invention is by appended power
Profit requires rather than described above limits, it is intended that all in the implication and scope of the equivalency of claim by falling
Change is included in the present invention.
Moreover, it will be appreciated that although the present specification is described in terms of embodiments, not each embodiment is only wrapped
Containing an independent technical scheme, this narrating mode of specification is only that those skilled in the art should for clarity
Using specification as an entirety, the technical solutions in the various embodiments may also be suitably combined, forms those skilled in the art
It is appreciated that other embodiment.
Claims (9)
1. the magnetic microparticle chemiluminescence detection kit and its composition of a kind of intravascular ErbB1, its feature exist
In：Including coupling have the magnetic particle suspension of intravascular ErbB1 capture antibody, acridinium ester label it is intravascular
ErbB1 detection antibody, calibration object, chemiluminescence preexciting liquid A, chemiluminescence exciting liquid B and cleaning fluid.
A kind of 2. magnetic microparticle chemiluminescence detection reagent of intravascular ErbB1 according to claim 1
Box and its composition, it is characterised in that：The kernel of described magnetic bead is ferroso-ferric oxide or di-iron trioxide, and the surface of magnetic particle is repaiied
Decorations group is one or more activity functional groups, and including but not limited to carboxyl, amino, p-toluenesulfonyl or strepto- is affine
Element-biotin.
A kind of 3. magnetic microparticle chemiluminescence detection reagent of intravascular ErbB1 according to claim 1
Box and its composition, it is characterised in that：Described magnetic particle can be directly even with intravascular ErbB1 capture antibody
Connection, or magnetic particle and Streptavidin are coupled, while it is anti-using the intravascular ErbB1 capture of biotin labeling
Body.
A kind of 4. magnetic microparticle chemiluminescence detection reagent of intravascular ErbB1 according to claim 1
Box and its composition, it is characterised in that：Described chemiluminescent labels are acridinium ester, such as NSP-DMAE-NHS, NSP-SA-NHS
Deng.
A kind of 5. magnetic microparticle chemiluminescence detection reagent of intravascular ErbB1 according to claim 1
Box and its composition, it is characterised in that：Described acridinium ester label is intravascular ErbB1 detection antibody.
A kind of 6. magnetic microparticle chemiluminescence detection reagent of intravascular ErbB1 according to claim 1
Box and its composition, it is characterised in that：Described capture antibody is monoclonal antibody with detection antibody.
A kind of 7. magnetic microparticle chemiluminescence detection reagent of intravascular ErbB1 according to claim 1
Box and its composition, it is characterised in that：Described calibration object is using the buffer solution containing BSA as matrix, adds the life of blood vessel endepidermis
The calibration object solution for the series concentration gradient that the configuration of the sterling of growth factor receptor body 1 forms.
A kind of 8. magnetic microparticle chemiluminescence detection reagent of intravascular ErbB1 according to claim 1
Box and its composition, it is characterised in that：Described chemiluminescence preexciting liquid A is by H2O2And HNO3Mixed liquor composition, chemiluminescence
Exciting liquid B is made up of Triton X-100 and NaOH mixed liquor.
A kind of 9. magnetic microparticle chemiluminescence detection reagent of intravascular ErbB1 according to claim 1
Box and its composition, comprise the following steps：A certain amount of sample to be tested is added in reaction cup, the coupling of magnetic particle is added and suspends
Liquid, mix, 37 DEG C of incubations, add cleaning fluid and remove supernatant；Acridinium ester label is added according to certain ratio afterwards, 37 DEG C incubate
Educate, add cleaning fluid and remove supernatant, add chemiluminescence preexciting liquid A and exciting liquid B, determine corresponding luminous intensity RLU/s.
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| CN111175516A (en) * | 2020-01-06 | 2020-05-19 | 潍坊市康华生物技术有限公司 | Vascular endothelial growth factor detection kit and preparation method thereof |
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| CN117825708A (en) * | 2024-03-04 | 2024-04-05 | 宁波美康盛德生物科技有限公司 | sFlt-1 detection kit |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101226200A (en) * | 2007-01-19 | 2008-07-23 | 天津天美生物技术有限公司 | Chemiluminescence immunity analysis detecting myocardium calcium protein T hypersensitization method for acridine ester and alkaline phosphatase |
| WO2010025414A3 (en) * | 2008-08-29 | 2010-05-14 | Genentech, Inc. | Diagnostics and treatments for vegf-independent tumors |
| US20110166166A1 (en) * | 2007-01-31 | 2011-07-07 | Henkin Robert I | Methods for detection of biological substances |
| CN105259163A (en) * | 2015-10-26 | 2016-01-20 | 深圳华迈兴微医疗科技有限公司 | Magnetic particle direct chemiluminiscence microfluidic chip for whole blood sample testing |
| CN106855572A (en) * | 2016-06-30 | 2017-06-16 | 深圳市亚辉龙生物科技股份有限公司 | A kind of gastrin-releasing peptide precursor chemiluminescence immune detection reagent kit and preparation method thereof |
-
2017
- 2017-11-03 CN CN201711068672.6A patent/CN107807241A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101226200A (en) * | 2007-01-19 | 2008-07-23 | 天津天美生物技术有限公司 | Chemiluminescence immunity analysis detecting myocardium calcium protein T hypersensitization method for acridine ester and alkaline phosphatase |
| US20110166166A1 (en) * | 2007-01-31 | 2011-07-07 | Henkin Robert I | Methods for detection of biological substances |
| WO2010025414A3 (en) * | 2008-08-29 | 2010-05-14 | Genentech, Inc. | Diagnostics and treatments for vegf-independent tumors |
| CN105259163A (en) * | 2015-10-26 | 2016-01-20 | 深圳华迈兴微医疗科技有限公司 | Magnetic particle direct chemiluminiscence microfluidic chip for whole blood sample testing |
| CN106855572A (en) * | 2016-06-30 | 2017-06-16 | 深圳市亚辉龙生物科技股份有限公司 | A kind of gastrin-releasing peptide precursor chemiluminescence immune detection reagent kit and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 毛光华: "血清及组织中VEGF，VEGFR1与VEGFR2在肺癌中的表达及其相互间的关系", 《万方学位论文数据库》 * |
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| CN109709336A (en) * | 2018-12-26 | 2019-05-03 | 张嘉庆 | A kind of detection kit and detection method of vascular endothelial growth factor |
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| CN117003878A (en) * | 2023-09-28 | 2023-11-07 | 南京佰抗生物科技有限公司 | Monoclonal antibody composition for resisting sFLT-1 protein and application thereof |
| CN117003878B (en) * | 2023-09-28 | 2023-12-05 | 南京佰抗生物科技有限公司 | Monoclonal antibody for resisting sFLT-1 protein and application thereof |
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