CN110082521A - A kind of quantum dot-preparation method of Creatine Kinase MB antibody immune complex and the preparation method of test strips - Google Patents
A kind of quantum dot-preparation method of Creatine Kinase MB antibody immune complex and the preparation method of test strips Download PDFInfo
- Publication number
- CN110082521A CN110082521A CN201910292989.0A CN201910292989A CN110082521A CN 110082521 A CN110082521 A CN 110082521A CN 201910292989 A CN201910292989 A CN 201910292989A CN 110082521 A CN110082521 A CN 110082521A
- Authority
- CN
- China
- Prior art keywords
- quantum dot
- creatine kinase
- antibody
- preparation
- immune complex
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
Abstract
The present invention relates to a kind of quantum dot-Creatine Kinase MB antibody immune complex preparation methods, it reacts quantum dot with 1- ethyl -3- (3- DimethylAminopropyl) carbodiimide hydrochloride and n-hydroxysuccinimide or its thio object, controls quantum dot, the molar ratio of 1- ethyl -3- (3- DimethylAminopropyl) carbodiimide hydrochloride, n-hydroxysuccinimide or its thio object is 1:4000 ~ 10000:5000 ~ 10000；The pH for adjusting reaction solution is 6 ~ 8；Creatine Kinase MB antibody is added into reaction solution, the molar ratio for controlling quantum dot and Creatine Kinase MB antibody is 1:10 ~ 18.Coupling effect of the present invention is good, and preparation method is simple, low in cost.The standard curve of test strips produced by the present invention is good, and detection sensitivity is high, can be realized the accurate quantitative analysis detection of Creatine Kinase MB antigen.
Description
Technical field
The invention belongs to biomarker fields, and in particular to a kind of quantum dot-Creatine Kinase MB antibody mediated immunity is multiple
Close the preparation method of object and the preparation method of test strips.
Background technique
Quantum dot (Quantum dots, QDs) is mainly by II B race ~ VI A race element (such as CdSe, CdTe, CdS, ZnSe
Deng) or III A race ~ V A race element (such as InP, InAs) composition semiconductor nanoparticle, three dimensions are in 100nm
Hereinafter, appearance is just like a pointing object.The quanta point biological compatibility of synthesis in water is good, is readily incorporated functional groups.Quantum
Point has special fluorescent characteristic compared with the organic conventional fluorescent material of tradition: 1) emission spectrum can be by changing size
It is controlled with composition, Color tunable is, it can be achieved that quantum dot polychrome of the same race marks；2) organic dyestuff fluorescent molecule exciting light is single,
Emission spectrum is wide, there is a hangover, and quantum dot is due to its special quantum confined effect, exciting light spectrum width, and emission spectrum is narrow and right
Claim, no long wave hangover；3) biggish Stokes shift avoids excitation spectrum overlapping with emission spectrum, is conducive to fluorescence spectrum
Detection；4) photochemical stability is high, and resistance to photobleaching can be subjected to repeated multiple times excitation；5) fluorescence efficiency is high；6) bio-compatible
Property is good, after various chemical modifications, can carry out specific connection；7) fluorescence lifetime is long, when exciting light close several nanoseconds with
Afterwards, quantum dot fluorescence still has, and can get the fluorescence signal without background interference.
And quantum dot is as novel nano material, the preparation method of biological fluorescent labelling is divided into non-covalent linking and altogether
Valence connection.Wherein being not covalently linked mainly has Electrostatic Absorption, the connection of specific biological target, streptomysin-biotin connection etc.；Altogether
Valence connection includes quantum dot surface functional group carboxyl, amino, hydroxyl etc., is divided respectively with biology under the activation of different activator
Amino, sulfydryl of son etc. are covalently attached.Wherein quantum dot surface carboxyl connect with biomolecule amino covalence and is most widely used.
But prepare that quantum dot-capture antibody complex method is different, and purification process needs the instrument of Large expensive at present
It completes, limits quantum dot-capture antibody complex study condition, coupling efficiency can not be calculated, limit quantum dot in life
Application in terms of substance markers.Secondly, the not no verification method of Applied economy, affects the detection performance of Related product.
It is cardiac muscle containing the more organ of Creatine Kinase MB (CK-MB), CK-MB is the early diagnosis of myocarditis disease
Important indicator, especially when diagnosing transmural type myocardial infarction, have high sensibility and specificity.In Healthy Human Serum
The content of CK-MB, hereinafter, if CK-MB content significantly increases in serum, illustrates myocardial damage 5%.In facing for heart infarction context of detection
Bed meaning are as follows: its content of 4 h increases after the onset of heart infarction, peaks in 16-24 h, and 3-4 d restore normal.If CK-MB exists
High level is still kept after acute myocardial infarction AMI, shows that myocardial necrosis is also carrying out；If increasing again after restoring normal, illustrate former stalk
Dead position extension or new infarct location occur, therefore CK-MB can be used as the early diagnosis marker of AMI.Immune test paper
The detection of item belongs to instant detection, can complete the quantitative detection to Troponin I in a short period of time, to meet clinical inspection
Survey the requirement of Creatine Kinase MB.
CN104280545A disclose a kind of quantum dot-labeled test strips for synchronizing quantitative Applications of Cardiac Markers multi objective and its
Method, still, inventor has found that being unable to get the mark of corresponding Applications of Cardiac Markers only with the content of the patent disclosure
Directrix curve, to cannot achieve the detection of corresponding Applications of Cardiac Markers.
Summary of the invention
Technical problem to be solved by the invention is to provide a kind of good quantum dot-creatine kinases of calibration curve coefficient correlation
The preparation method of isodynamic enzyme MB antibody immune complex and the preparation method of test strips.
In order to solve the above technical problems, the present invention adopts the following technical scheme that:
It is an object of the present invention to provide a kind of quantum dot-Creatine Kinase MB antibody immune complex preparation sides
Method, characterized by the following steps:
(1), make quantum dot and 1- ethyl -3- (3- DimethylAminopropyl) carbodiimide hydrochloride and n-hydroxysuccinimide
Or its thio object is reacted, and the quantum dot, described 1- ethyl -3- (3- DimethylAminopropyl) carbodiimides are controlled
The molar ratio of hydrochloride, the n-hydroxysuccinimide or its thio object is 1:4000 ~ 10000:5000 ~ 10000；
(2), the pH of set-up procedure (1) treated reaction solution is 6 ~ 8；
(3), quantum dot-flesh that Creatine Kinase MB antibody is made described is added into step (2) treated reaction solution
Acid kinase isodynamic enzyme MB antibody immune complex controls the throwing of the quantum dot and the Creatine Kinase MB antibody
Material molar ratio is 1:10 ~ 18.
In the present invention, the surface of the quantum dot is carboxyl modified, and the quantum dot can be hud typed or non-core
Shell type quantum point.
Preferably, in step (1), the average grain diameter of the quantum dot is 10 ~ 30nm, launch wavelength range be 560nm ~
650nm。
Preferably, in step (1), the quantum dot, 1- ethyl -3- (3- DimethylAminopropyl) carbonization two are sub-
The molar ratio of amine hydrochlorate, the n-hydroxysuccinimide or its thio object be 1:4000 ~ 6000:8000 ~
10000。
Preferably, the reaction condition of step (1) is ice-bath ultrasonic, and the reaction time is 20 ~ 40min.
According to a specific and preferred embodiment, the specific embodiment of step (1) are as follows: it is 5 ~ 6 that quantum dot, which is dissolved in pH,
In the borate buffer solution of 0.005 ~ 0.015M, it is sub- that 1- ethyl -3- (3- DimethylAminopropyl) carbonization two is then added
Amine hydrochlorate and the n-hydroxysuccinimide or its thio object carry out priming reaction.
Preferably, step (1) after the reaction was completed, removes excessive 1- ethyl -3- (3- dimethyl amine third using ultrafiltration
Base) carbodiimide hydrochloride and n-hydroxysuccinimide or its thio object, the borate buffer for being then 6 ~ 8 by addition pH
The pH of liquid adjustment reaction solution.
Preferably, in step (3), the molar ratio of the quantum dot and the Creatine Kinase MB antibody
For 1:13 ~ 16.The present invention passes through the dosage of strict control antibody and quantum dot, ensure that the joint efficiency of quantum dot and antibody.
Preferably, the reaction condition of step (3) is ice-bath ultrasonic, and the reaction time is 2 ~ 5h.
It is a further object to provide a kind of preparation methods of test strips, include the following steps:
(1) using treatment fluid processing sample pad and bonding pad；
(2) using quantum dot-Creatine Kinase MB antibody immune complex processing step (1) made from above-mentioned preparation method
Treated bonding pad；
(3) detection is coated on nitrocellulose filter using Creatine Kinase MB antibody and sheep anti-mouse igg polyclonal antibody
Line and nature controlling line, then using the nitrocellulose filter after confining liquid closing coating；
(4) by step (1) treated sample pad, step (2) treated bonding pad, step (3) treated cellulose nitrate
Plain film, water absorption pad and carrier stack gradually, and the test strips are made.
Preferably, in step (1), based on mass fraction, the treatment fluid be containing 1 ~ 5% bovine serum albumin(BSA), 1 ~
3% polysorbas20,0.5 ~ 1.5% NaCl, 1 ~ 5% sucrose and 1 ~ 3% polyethylene glycol 2000 pH be 7 ~ 9 borate buffer
Liquid；In step (3), based on mass fraction, the confining liquid is the bovine serum albumin(BSA) for being 0.5 ~ 2% containing mass fraction
The borate buffer solution that pH value is 7 ~ 9.
Preferably, in step (3), use Creatine Kinase MB antibody-solutions that concentration is 0.5 ~ 2mg/ml with 0.5 ~
Detection line is made in the scribing line speed of 1 μ l/cm on the nitrocellulose filter, uses concentration for the goat-anti of 0.5 ~ 1 mg/ml
Nature controlling line is made with the scribing line speed of 0.5 ~ 1 μ l/cm in mouse IgG Anti-TNF-α liquid solution on nitrocellulose filter.
It is further preferred that the Creatine Kinase MB antibody-solutions and the sheep anti-mouse igg Anti-TNF-α
Liquid solution dilutes to obtain using borate buffer.
It is further preferable that the borate buffer of Creatine Kinase MB antibody described in dilution is pH6.5, ion is strong
Degree: the borate buffer of 0.05M.
It is further preferable that the borate buffer of sheep anti-mouse igg polyclonal antibody described in dilution is pH5.5, ionic strength:
The borate buffer of 0.01M.
Further, front of the detection line in the immunochromatography direction of nature controlling line when described stroke of film.
Further, detection line and nature controlling line are close to water absorption pad side when described stroke of film, and purpose is when reserving enough
Between allow antibody antigen carry out specific reaction.
Due to the implementation of above technical scheme, the invention has the following advantages over the prior art:
The coupling effect of quantum dot of the invention and Creatine Kinase MB antibody is good, and preparation method is simple, low in cost.This
The standard curve for inventing test strips obtained is good, and related coefficient is greater than 0.99, and detection sensitivity is high, and it is same to can be realized creatine kinase
The accurate quantitative analysis of work enzyme MB antigen detects.
Detailed description of the invention
Fig. 1 is quantum dot-Creatine Kinase MB antibody of quantum dot control (left-hand bar band) and the embodiment of the present invention 1
The agarose gel electrophoresis figure of immune complex (right side band)；
Fig. 2 is in coupling reaction using quantum dot-Creatine Kinase MB antibody immune complex made from different pH value
Gel electrophoresis figure；
Fig. 3 is multiple using quantum dot-Creatine Kinase MB antibody mediated immunity made from different activator levels in coupling reaction
Close the gel electrophoresis figure of object；
Fig. 4 is compound using quantum dot-Creatine Kinase MB antibody mediated immunity made from different antibodies dosage in coupling reaction
The gel electrophoresis figure of object；
Fig. 5 is quantum dot-Creatine Kinase MB antibody immune complex partial size of quantum dot control and the embodiment of the present invention 1
Size detection figure；
Wherein, control of the Fig. 1 into Fig. 5 is the result that the quantum dot being directly commercially available is detected；
Fig. 6 is that quantum dot compares the test strips with the embodiment of the present invention 1 in loading gradient Creatine Kinase MB antigen concentration
Ultraviolet figure afterwards；
Fig. 7 is that quantum dot compares the test strips with the embodiment of the present invention 1 in loading gradient Creatine Kinase MB antigen concentration
When obtained canonical plotting.
Specific embodiment
The present invention will be further described in detail combined with specific embodiments below, but the present invention is not limited to following implementations
Example.Implementation condition used in the examples can do further adjustment according to specifically used different requirements, the implementation being not specified
Condition is the normal condition in the industry, the commercially available acquisition of reagent in the present invention.In following, unless otherwise specified, " % " is equal
For mass percentage.
Embodiment 1
A kind of preparation of quantum dot-Creatine Kinase MB antibody immune complex:
5ul quantum dot (average grain diameter is 18 nm, and emission peak is 625nm ± 5nm, concentration 8uM, is purchased from Wuhan Ka source) is molten
In 500ul, pH 5.5 in 0.01M borate buffer solution, is vortexed and mixes 30s.Now prepare 0.01M 1- ethyl -3- (3- dimethyl
Amine propyl) carbodiimide hydrochloride and 0.01M N- hydroxy thiosuccinimide, solvent is pH 5.5,0.01M borate
Buffer.20 ul, 0.01M 1- ethyl -3- (3- DimethylAminopropyl) carbodiimides hydrochloric acid are added in quantum dot solution
Salt is vortexed and mixes, and 40ul is added after 5min, and 0.01M N- hydroxy thiosuccinimide is vortexed and mixes.Ice-bath ultrasonic 30min.
After the completion of activation, priming reaction liquid is taken out, is placed in super filter tube (100kd), is centrifuged in refrigerated centrifuge, revolving speed 3500rpm,
1ml is added after the completion in 7min in reaction solution, and pH 6.5,0.01M borate buffer solution, repeated centrifugation is primary, takes out reaction
Liquid.7ul Creatine Kinase MB capture antibody (anti-CK-MB-1363,7.3mg/ml) is added into reaction solution, is vortexed
It mixes, ice-bath ultrasonic 3h is to get quantum dot-Creatine Kinase MB antibody immune complex.
The quantum dot-Creatine Kinase MB antibody immune complex agarose gel electrophoresis figure is as shown in Figure 1, grain
Diameter size detection figure is as shown in Figure 5, wherein the average grain diameter of quantum dot is 18nm, quantum dot-Creatine Kinase MB antibody
The average grain diameter of immune complex is 86nm, from Fig. 1 and Fig. 5 as it can be seen that embodiment 1 successfully to have obtained quantum dot-creatine kinase same
Work enzyme MB antibody immune complex.
The preparation of test strips:
(1) sample pad and bonding pad are respectively placed in treatment fluid (containing 1% bovine serum albumin(BSA), 1% polysorbas20,0.85%
The borate buffer of the pH8.5 of NaCl, 1% sucrose and 2% polyethylene glycol 2000) in, it impregnates 2 hours, then in 37 DEG C of perseverances
It is taken out after drying in warm incubator spare.
(2) quantum dot-Creatine Kinase MB made from the above method is sprayed on step (1) treated bonding pad
Antibody immune complex and drying, control spray rate are 0.8 μ L/cm.
(3) boric acid for Creatine Kinase MB antibody (HyTest company) being dissolved in pH6.5, ionic strength: 0.05M is slow
It is configured to the Creatine Kinase MB antibody-solutions that concentration is 1mg/ml in fliud flushing, is then existed with the scribing line speed of 0.5 μ L/cm
Detection line is made in drying of crossing on nitrocellulose filter；Sheep anti-mouse igg polyclonal antibody (HyTest) is dissolved in pH5.5, ion
Intensity: it is configured to the mouse that concentration is 0.5 mg/ml in the borate buffer of 0.01M and resists how anti-solution, then with 0.5 μ l/cm's
Scribing line speed is crossed on nitrocellulose filter dries obtained nature controlling line.
(4) use containing mass fraction for 1% bovine serum albumin(BSA), pH value be 8.5 borate buffer to draw have detection
The nitrocellulose filter of line and nature controlling line impregnates two hours, after dry in 37 DEG C of constant incubators take out.
(5) by step (1) treated sample pad, step (2) treated bonding pad, step (4) treated nitric acid
Cellulose membrane, water absorption pad and carrier stack gradually, and test strips are made.
(6) the Creatine Kinase MB antigenic solution containing various concentration is added drop-wise to the sample pad surface of test strips,
Obtained ultraviolet figure is shown in that Fig. 6, canonical plotting are shown in Fig. 7.
Embodiment 2
Substantially same as Example 1, the difference is that: preparing, quantum dot-Creatine Kinase MB antibody mediated immunity is compound
When object, the borate buffer solution that different pH are respectively adopted adjusts the pH of reaction solution after activation, and agarose gel electrophoresis figure is shown in Fig. 2,
Wherein, 1 is the band compareed.Quantum dot-Creatine Kinase MB antibody made from 2 borate buffer solution for pH5.5 is exempted from
The band of epidemic disease compound, 3 be that quantum dot-Creatine Kinase MB antibody mediated immunity made from the borate buffer solution of pH6.5 is multiple
Close the band of object, quantum dot-Creatine Kinase MB antibody immune complex made from 4 borate buffer solution for pH7.5
Band, 5 for pH8.5 borate buffer solution made from quantum dot-Creatine Kinase MB antibody immune complex item
Band, 6 be quantum dot-Creatine Kinase MB antibody immune complex band made from the borate buffer solution of pH9.5.
It analyzes since the isoelectric point of different antibody is different, in different pH, the electrification of antibody is different, affects antibody
With the coupling effect of quantum dot, quantum dot is coupled with different antibodies, that is, has different best coupling pH.
Outside test strips made from borate buffer solution when condition is pH6.5 and 7.5, examination obtained under the conditions of other pH
Paper slip is unable to get standard curve, also, the related coefficient of the standard curve of test strips obtained is inferior under the conditions of pH7.5
The related coefficient of the standard curve of test strips obtained under the conditions of pH6.5.
Embodiment 3
Substantially same as Example 1, the difference is that: preparing, quantum dot-Creatine Kinase MB antibody mediated immunity is compound
When object, the quantum dot (QDs) and 1- ethyl -3- (3- DimethylAminopropyl) carbodiimides salt of different mol ratio example are added respectively
Hydrochlorate (EDC), agarose gel electrophoresis figure is shown in Fig. 3, wherein 1 is carbonized for quantum dot and 1- ethyl -3- (3- DimethylAminopropyl)
The molar ratio of diimmonium salt hydrochlorate is the band of 1:200, and 2 are carbonized for quantum dot and 1- ethyl -3- (3- DimethylAminopropyl)
The molar ratio of diimmonium salt hydrochlorate is the band of 1:1000, and 3 are carbonized for quantum dot and 1- ethyl -3- (3- DimethylAminopropyl)
The molar ratio of diimmonium salt hydrochlorate is the band of 1:5000, and 4 are carbonized for quantum dot and 1- ethyl -3- (3- DimethylAminopropyl)
The molar ratio of diimmonium salt hydrochlorate is the band of 1:25000, and 5 be quantum dot and 1- ethyl -3- (3- DimethylAminopropyl) carbon
The molar ratio for changing diimmonium salt hydrochlorate is the band of 1:50000.
Analysis is since QDs is different from the molar ratio of EDC, the activation degree difference of QDs surface carboxyl groups, then with ammonia in antibody
The case where base reacts is different, causes to be coupled success rate difference.
Embodiment 4
Substantially same as Example 1, the difference is that: preparing, quantum dot-Creatine Kinase MB antibody mediated immunity is compound
When object, the quantum dot and Creatine Kinase MB for adding different mol ratio example respectively capture antibody, agarose gel electrophoresis figure
See Fig. 4, wherein 1 is that quantum dot and Creatine Kinase MB capture the band that the molar ratio of antibody is 1:2, and 2 be quantum dot
The band that molar ratio with Creatine Kinase MB capture antibody is 1:6,3 catch for quantum dot with Creatine Kinase MB
The molar ratio for obtaining antibody is the band of 1:10, and 4 be that quantum dot and Creatine Kinase MB capture the molar ratio of antibody and be
The band of 1:14,5 be that quantum dot and Creatine Kinase MB capture the band that the molar ratio of antibody is 1:18.
The present invention is described in detail above, its object is to allow the personage for being familiar with this field technology that can understand this
The content of invention is simultaneously implemented, and it is not intended to limit the scope of the present invention, all Spirit Essence institutes according to the present invention
The equivalent change or modification of work, should be covered by the scope of protection of the present invention.
Claims (10)
1. a kind of quantum dot-Creatine Kinase MB antibody immune complex preparation method, it is characterised in that: including as follows
Step:
(1), make quantum dot and 1- ethyl -3- (3- DimethylAminopropyl) carbodiimide hydrochloride and n-hydroxysuccinimide
Or its thio object is reacted, and the quantum dot, described 1- ethyl -3- (3- DimethylAminopropyl) carbodiimides are controlled
The molar ratio of hydrochloride, the n-hydroxysuccinimide or its thio object is 1:4000 ~ 10000:5000 ~ 10000；
(2), the pH of set-up procedure (1) treated reaction solution is 6 ~ 8；
(3), quantum dot-flesh that Creatine Kinase MB antibody is made described is added into step (2) treated reaction solution
Acid kinase isodynamic enzyme MB antibody immune complex controls the throwing of the quantum dot and the Creatine Kinase MB antibody
Material molar ratio is 1:10 ~ 18.
2. quantum dot according to claim 1-Creatine Kinase MB antibody immune complex preparation method, special
Sign is: in step (1), the average grain diameter of the quantum dot is 10 ~ 30nm, and launch wavelength range is 560nm ~ 650nm.
3. quantum dot according to claim 1-Creatine Kinase MB antibody immune complex preparation method, special
Sign is: in step (1), the quantum dot, described 1- ethyl -3- (3- DimethylAminopropyl) the carbodiimides hydrochloric acid
The molar ratio of salt, the n-hydroxysuccinimide or its thio object is 1:4000 ~ 6000:8000 ~ 10000.
4. quantum dot according to claim 1-Creatine Kinase MB antibody immune complex preparation method, special
Sign is: the reaction condition of step (1) is ice-bath ultrasonic, and the reaction time is 20 ~ 40min.
5. quantum dot according to claim 1-Creatine Kinase MB antibody immune complex preparation method, special
Sign is: step (1) removes excessive 1- ethyl -3- (3- DimethylAminopropyl) carbonization two after the reaction was completed, using ultrafiltration
Then inferior amine salt hydrochlorate and n-hydroxysuccinimide or its thio object are adjusted anti-by the borate buffer that addition pH is 6 ~ 8
Answer the pH of liquid.
6. quantum dot according to claim 1-Creatine Kinase MB antibody immune complex preparation method, special
Sign is: in step (3), the molar ratio of the quantum dot and the Creatine Kinase MB antibody be 1:13 ~
16。
7. quantum dot according to claim 1-Creatine Kinase MB antibody immune complex preparation method, special
Sign is: the reaction condition of step (3) is ice-bath ultrasonic, and the reaction time is 2 ~ 5h.
8. a kind of preparation method of test strips, characterized by the following steps:
(1) using treatment fluid processing sample pad and bonding pad；
(2) using quantum dot-Creatine Kinase MB made from the preparation method as described in any one of claims 1 to 7
Antibody immune complex processing step (1) treated bonding pad；
(3) detection is coated on nitrocellulose filter using Creatine Kinase MB antibody and sheep anti-mouse igg polyclonal antibody
Line and nature controlling line, then using the nitrocellulose filter after confining liquid closing coating；
(4) by step (1) treated sample pad, step (2) treated bonding pad, step (3) treated cellulose nitrate
Plain film, water absorption pad and carrier stack gradually, and the test strips are made.
9. preparation method according to claim 8, it is characterised in that: in step (1), based on mass fraction, the place
Manage liquid be containing 1 ~ 5% bovine serum albumin(BSA), 1 ~ 3% polysorbas20,0.5 ~ 1.5% NaCl, 1 ~ 5% sucrose and 1 ~ 3% it is poly-
The borate buffer that the pH of ethylene glycol 2000 is 7 ~ 9；In step (3), based on mass fraction, the confining liquid is to contain quality
The borate buffer solution that the pH value for the bovine serum albumin(BSA) that score is 0.5 ~ 2% is 7 ~ 9.
10. preparation method according to claim 8, it is characterised in that: in step (3), use concentration for 0.5 ~ 2mg/ml
Creatine Kinase MB antibody-solutions with the scribing line speed of 0.5 ~ 1 μ l/cm on the nitrocellulose filter be made inspection
Survey line, use concentration be 0.5 ~ 1 mg/ml sheep anti-mouse igg Anti-TNF-α liquid solution with the scribing line speed of 0.5 ~ 1 μ l/cm in nitre
Nature controlling line is made on acid cellulose film.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910292989.0A CN110082521A (en) | 2019-04-12 | 2019-04-12 | A kind of quantum dot-preparation method of Creatine Kinase MB antibody immune complex and the preparation method of test strips |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910292989.0A CN110082521A (en) | 2019-04-12 | 2019-04-12 | A kind of quantum dot-preparation method of Creatine Kinase MB antibody immune complex and the preparation method of test strips |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110082521A true CN110082521A (en) | 2019-08-02 |
Family
ID=67414862
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910292989.0A Pending CN110082521A (en) | 2019-04-12 | 2019-04-12 | A kind of quantum dot-preparation method of Creatine Kinase MB antibody immune complex and the preparation method of test strips |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110082521A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111879921A (en) * | 2020-06-22 | 2020-11-03 | 武汉生之源生物科技股份有限公司 | Fluorescent microsphere of coupled antibody and preparation method and application thereof |
| CN111896749A (en) * | 2020-08-07 | 2020-11-06 | 杭州都林生物科技有限公司 | Bovine lactoferrin double-antibody sandwich colloidal gold immunochromatographic test strip, preparation method, kit and detection method |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102520192A (en) * | 2011-12-29 | 2012-06-27 | 深圳康美生物科技股份有限公司 | Fluorescence immunochromatographic assay and kit for quantitative detection of troponin I/creatine kinase isoenzyme/myohemoglobin |
| CN102520173A (en) * | 2011-12-29 | 2012-06-27 | 深圳康美生物科技股份有限公司 | Fluorescence immunochromatographic assay and kit for quantitative detection of creatine kinase isoenzyme (CK-MB) |
| CN102520194A (en) * | 2011-12-29 | 2012-06-27 | 深圳康美生物科技股份有限公司 | Fluorescence immunochromatographic assay method for quantitatively detecting heart fatty acid binding protein and kit for quantitatively detecting same |
| CN105842463A (en) * | 2016-05-17 | 2016-08-10 | 北京美康基因科学股份有限公司 | Kit for myocardial function assay and preparation method thereof |
| CN108918882A (en) * | 2018-05-17 | 2018-11-30 | 西安交通大学苏州研究院 | A kind of preparation method of the hs-CRP immuno-chromatographic test paper strip based on quantum dot |
| CN108918858A (en) * | 2018-05-17 | 2018-11-30 | 西安交通大学苏州研究院 | A kind of preparation method of quantum dot-antibody immune complex |
| CN108982828A (en) * | 2018-06-28 | 2018-12-11 | 长春万成生物电子工程有限公司 | A kind of quantum dot examines the preparation and application of test paper fastly |
-
2019
- 2019-04-12 CN CN201910292989.0A patent/CN110082521A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102520192A (en) * | 2011-12-29 | 2012-06-27 | 深圳康美生物科技股份有限公司 | Fluorescence immunochromatographic assay and kit for quantitative detection of troponin I/creatine kinase isoenzyme/myohemoglobin |
| CN102520173A (en) * | 2011-12-29 | 2012-06-27 | 深圳康美生物科技股份有限公司 | Fluorescence immunochromatographic assay and kit for quantitative detection of creatine kinase isoenzyme (CK-MB) |
| CN102520194A (en) * | 2011-12-29 | 2012-06-27 | 深圳康美生物科技股份有限公司 | Fluorescence immunochromatographic assay method for quantitatively detecting heart fatty acid binding protein and kit for quantitatively detecting same |
| CN105842463A (en) * | 2016-05-17 | 2016-08-10 | 北京美康基因科学股份有限公司 | Kit for myocardial function assay and preparation method thereof |
| CN108918882A (en) * | 2018-05-17 | 2018-11-30 | 西安交通大学苏州研究院 | A kind of preparation method of the hs-CRP immuno-chromatographic test paper strip based on quantum dot |
| CN108918858A (en) * | 2018-05-17 | 2018-11-30 | 西安交通大学苏州研究院 | A kind of preparation method of quantum dot-antibody immune complex |
| CN108982828A (en) * | 2018-06-28 | 2018-12-11 | 长春万成生物电子工程有限公司 | A kind of quantum dot examines the preparation and application of test paper fastly |
Non-Patent Citations (2)
| Title |
|---|
| JING ZHANG 等: ""Aptamer-based fluorometric lateral flow assay for creatine kinase MB"", 《MICROCHIMICA ACTA》 * |
| 褚春旭 等: ""心肌损伤标志物肌酸激酶同工酶荧光检测方法的建立"", 《长春理工大学学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111879921A (en) * | 2020-06-22 | 2020-11-03 | 武汉生之源生物科技股份有限公司 | Fluorescent microsphere of coupled antibody and preparation method and application thereof |
| CN111896749A (en) * | 2020-08-07 | 2020-11-06 | 杭州都林生物科技有限公司 | Bovine lactoferrin double-antibody sandwich colloidal gold immunochromatographic test strip, preparation method, kit and detection method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Mahmoud et al. | Advanced procedures for labeling of antibodies with quantum dots | |
| JP6260541B2 (en) | An immunoassay to reduce the effects of impurities | |
| CN111308103B (en) | Cardiopulmonary quintuplet detection kit, rare earth nano fluorescence detection card and detection method thereof | |
| US11668706B2 (en) | Near-infrared II polymer fluorescent microsphere and method for preparing same | |
| Filbrun et al. | Chemical modification of antibodies enables the formation of stable antibody–gold nanoparticle conjugates for biosensing | |
| Zhang et al. | Improving detection sensitivity by oriented bioconjugation of antibodies to quantum dots with a flexible spacer arm for immunoassay | |
| CN110082521A (en) | A kind of quantum dot-preparation method of Creatine Kinase MB antibody immune complex and the preparation method of test strips | |
| Zhang et al. | Glucose oxidase-directed, instant synthesis of Mn-doped ZnS quantum dots in neutral media with retained enzymatic activity: mechanistic study and biosensing application | |
| CN110520734A (en) | The method for generating the noise in number measurement for reducing signal | |
| CN108181287A (en) | A kind of magnetic bead time-resolved fluoroimmunoassay quantitatively detects H-FABP kits | |
| Nguyen et al. | Ultrasensitive biogenic amine sensor using an enhanced multiple nanoarray chip based on competitive reactions in an evanescent field | |
| CN110082522A (en) | A kind of quantum dot-preparation method of cardiac muscle troponin I antibody immune complex and the preparation method of test strips | |
| CN110082523A (en) | A kind of quantum dot-preparation method of myoglobins antibody immune complex and the preparation method of test strips | |
| JP3937020B2 (en) | Surface plasmon resonance antibody array sensor fabrication substrate and fabrication method thereof | |
| CN104777317B (en) | The preparation of a kind of gold nanoparticle probe and the application in tachysynthesis detects thereof | |
| CN210923728U (en) | Time-resolved fluorescence S-100 β protein detection card | |
| CN112114149A (en) | Application of receptor reagent in diagnosis of myocardial damage of main body | |
| CN112240936A (en) | Application of donor reagent in diagnosis of myocardial damage of main body | |
| JP2561134B2 (en) | Monoclonal antibody-derived substance used for elimination / suppression of non-specific reaction in immunoassay, production method thereof and use thereof | |
| CN106198999A (en) | The preparation method and applications of quantum dot β HCG monoclonal antibody conjugate | |
| CN113125721B (en) | Homogeneous detection kit for creatine kinase isozymes and application thereof | |
| CN113125703B (en) | Myoglobin homogeneous detection kit and application thereof | |
| RU2814452C1 (en) | Fluorescent hydrogel for detecting biological molecules | |
| CN113125705B (en) | Myoglobin homogeneous detection kit and application thereof | |
| CN113125700B (en) | Homogeneous detection kit for creatine kinase isozymes and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190802 |