CN105891194A - Anticardiolipin antibody IgG chemiluminescence immunoassay kit and preparation method thereof - Google Patents
Anticardiolipin antibody IgG chemiluminescence immunoassay kit and preparation method thereof Download PDFInfo
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- CN105891194A CN105891194A CN201610516757.5A CN201610516757A CN105891194A CN 105891194 A CN105891194 A CN 105891194A CN 201610516757 A CN201610516757 A CN 201610516757A CN 105891194 A CN105891194 A CN 105891194A
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Abstract
The invention discloses an anticardiolipin antibody IgG chemiluminescence immunoassay kit and a preparation method thereof. The anticardiolipin antibody IgG chemiluminescence immunoassay kit comprises an anticardiolipin antibody IgG monoclonal antibody coated carboxylated magnetic micro-particles and an anti-human IgG secondary antibody marked chemiluminescence marker. The anticardiolipin antibody IgG chemiluminescence immunoassay kit can take a full-automatic chemiluminescence immunity analyzer as a detection tool to finish detection of an anticardiolipin antibody IgG; an experiment of the anticardiolipin antibody IgG chemiluminescence immunoassay kit shows that the detection sensitivity reaches 10pg/mL; compared with a traditional anticardiolipin antibody IgG detection method, the sensitivity is improved by at least 10 times; the anticardiolipin antibody IgG chemiluminescence immunoassay kit has higher detection precision.
Description
Technical field
The present invention relates to vitro detection field, particularly relate to a kind of anticardiolipin antibody IgG chemiluminescence immunoassay
Detection kit and preparation method thereof.
Background technology
Cuorin is that the one separated from the cardiac muscle of cattle has antigenic phospholipid, and is named,
Its content in mammal flesh is the highest, and each Cardiolipin molecules comprises 4 undersaturated fatty acids, easily
Oxidation.Anticardiolipin antibody (anticardiolipin, ACL) is a kind of negative with band in platelet and endothelial cell membrane
The cuorin of electric charge is as the autoantibody of target antigen.See the Patients with SLE peace treaty of 50%
Other system autoimmune patients with abnormal (rheumatoid arthritis, scleroderma, the sjogren syndrome of 4-50%
Deng).This antibody is in close relations with thrombosis, platelet, spontaneous abortion or FDIU.Cardiolipin antibody
Can have IgA, IgG or IgM hypotype, what diagnostic value was the highest is high concentration IgG antibody, but a lot of patient's blood
IgA and IgM type cardiolipin antibody can be detected in Qing.Additionally, evidence suggests the anticardiolipin IgG of high concentration
Type antibody is relevant with thrombocytopenia height, and the resistance phospholipid IgM type antibody of high concentration and hemolytic are lean
Blood height is relevant.
The main method of Clinical detection anticardiolipin antibody IgG is enzyme linked immunosorbent assay, but the method exists
Following weak point:
(1) use 12 × 8 types, 6 × 8 types, 8 × 12 types or complete plate 96 hole Special micro porous plate as antigen bag
By apparatus and reaction vessel, 12 batches, 6 batches, 8 batches or imposite can only be divided in use and once make
With, it is impossible to carry out independent, the detection of single part；
(2) reagent type used by quantitative determination is more, and each detectable will contain with reagent bottle,
And it is required for changing imbibition nozzle when often using a kind of reagent to be filled into respectively in the micropore of microwell plate, not only
Reagent bottle kind is many, and the operation of filling reagent is the most loaded down with trivial details；
(3) the corresponding mark to detection information is lacked, can only be by checking the mark ability of test kit external packing box
Understand or know product batch number and the effect duration information of detectable, and the information known is in detection process
In uncontrolled, there is the biggest randomness；
(4) detectable is in open space during detection, easily causes the intersection between various reagent
Pollute and affect the accuracy of testing result；
(5) detection process uses manual operations, and the dosage of reagent or sample is not bery accurate, and operating process is extremely
Loaded down with trivial details and complicated, it is susceptible to bust, accuracy and the precision of testing result are poor；
(6) in the quantity configuration and use of detection project reagent set, it is item number × 48/96 person-portion, if
Need to detect 10 projects, then configuration and the use number of reagent must be 10 × 48/96 person-portions, if only portion
Sample needs to detect 10 different projects, it is also desirable to the reagent of configuration 10 × 48/96 person-portions, also exists not
The shortcoming of economical rationality.
Summary of the invention
Based on this, it is necessary to the anticardiolipin antibody IgG chemiluminescence providing a kind of detection sensitivity higher is exempted from
Epidemic disease detection kit and preparation method thereof.
A kind of anticardiolipin antibody IgG chemiluminescence immune detection reagent kit, including: anticardiolipin antibody IgG
The coated carboxylated magnetic particle of monoclonal antibody and the chemiluminescent labels of the anti-labelling of anti-human igg two.
In the coated carboxylated magnetic particle of described anticardiolipin antibody IgG monoclonal antibody, described anti-heart phosphorus
Fat IgG antibody monoclonal antibody is 1:25～35 with the ratio of described carboxylated magnetic particle.
In the chemiluminescent labels of the described anti-labelling of anti-human igg two, described anticardiolipin antibody IgG Dan Ke
Grand antibody is 50:1～10 with the ratio of described chemiluminescent labels.
The particle diameter of described carboxylated magnetic particle is 0.05 μm～1 μm.
Described chemiluminescent labels is luminol, different luminol, tris (bipyridine) ruthenium or acridinium ester.
Described anticardiolipin antibody IgG chemiluminescence immune detection reagent kit, also includes chemical luminous substrate
Liquid, described Chemoluminescent substrate includes A liquid and B liquid.
Described A liquid is H2O2Solution, described B liquid is NaOH solution.
Described anticardiolipin antibody IgG chemiluminescence immune detection reagent kit, also includes anticardiolipin antibody
IgG calibrates product.
Described anticardiolipin antibody IgG calibration product be concentration be respectively 0GPLU/mL, 10GPLU/mL,
Anticardiolipin antibody IgG of 20GPLU/mL, 50GPLU/mL, 100GPLU/mL and 200GPLU/mL
Solution.
A kind of preparation method of above-mentioned anticardiolipin antibody IgG chemiluminescence immune detection reagent kit, including
Following steps:
Taking the suspension of carboxylated magnetic particle, Magneto separate is resuspended with MES buffer after removing supernatant, then adds
Enter EDC aqueous solution, the surface carboxyl groups of the magnetic particle of activated carboxyl, it is subsequently added into anticardiolipin antibody IgG
Monoclonal antibody, suspendible 2h～10h under room temperature, Magneto separate is resuspended with Tris buffer after removing supernatant, obtains
The coated carboxylated magnetic particle of anticardiolipin antibody IgG monoclonal antibody；And
Take anticardiolipin antibody IgG monoclonal antibody, mix after adding carbonate buffer solution, being subsequently adding
Mixing after learning luminous marker, under room temperature, remove impurity after lucifuge reaction 1h～2h, obtains the anti-labelling of anti-human igg two
Chemiluminescent labels.
This anticardiolipin antibody IgG chemiluminescence immune detection reagent kit can be exempted from Full-automatic chemiluminescence
Epidemic disease analyser is detection instrument, completes the detection this anticardiolipin antibody IgGization of anticardiolipin antibody IgG
Learning electrochemiluminescent immunoassay detection kit, through experiment, its detection sensitivity reaches 0.2GPLU/mL, relative to biography
The detection method sensitivity of anticardiolipin antibody IgG of system at least improves 10 times, this anticardiolipin antibody
The accuracy of detection of IgG chemiluminescence immune detection reagent kit is higher.
Accompanying drawing explanation
Fig. 1 is the preparation side of the anticardiolipin antibody IgG chemiluminescence immune detection reagent kit of an embodiment
The flow chart of method；
Fig. 2 is the anticardiolipin antibody IgG canonical plotting that embodiment 3 obtains.
Detailed description of the invention
Understandable for enabling the above-mentioned purpose of the present invention, feature and advantage to become apparent from, below in conjunction with the accompanying drawings and
The detailed description of the invention of the present invention is described in detail by specific embodiment.Elaborate in the following description very
Many details are so that fully understanding the present invention.But the present invention can be described here to be much different from
Alternate manner is implemented, and those skilled in the art can do similar changing in the case of intension of the present invention
Entering, therefore the present invention is not limited by following public being embodied as.
The anticardiolipin antibody IgG chemiluminescence immune detection reagent kit of one embodiment, including: anti-heart phosphorus
The coated carboxylated magnetic particle of fat IgG antibody monoclonal antibody and the chemiluminescence of the anti-labelling of anti-human igg two
Label.
Preferably, in the coated carboxylated magnetic particle of anticardiolipin antibody IgG monoclonal antibody, anti-heart phosphorus
Fat IgG antibody monoclonal antibody is 1:25～35 with the ratio of carboxylated magnetic particle.
Preferably, in the chemiluminescent labels of the anti-labelling of anti-human igg two, anticardiolipin antibody IgG Dan Ke
Grand antibody is 50:1～10 with the ratio of chemiluminescent labels.
Preferably, the particle diameter of carboxylated magnetic particle is 0.05 μm～1 μm.
Chemiluminescent labels can be luminol, different luminol, tris (bipyridine) ruthenium or acridinium ester.Wherein,
Chemiluminescent labels is preferably acridinium ester.
In other examples, above-mentioned anticardiolipin antibody IgG chemiluminescence immune detection reagent kit also wraps
Include Chemoluminescent substrate.
Chemoluminescent substrate includes A liquid and B liquid.A liquid can be H2O2Solution, B liquid can be NaOH
Solution.
In the present embodiment, A liquid be concentration be the H of 0.1mol/L2O2Solution, B liquid be concentration be 0.25mol/L
NaOH solution.
In other examples, above-mentioned anticardiolipin antibody IgG chemiluminescence immune detection reagent kit also wraps
Include anticardiolipin antibody IgG calibration product.
Anticardiolipin antibody IgG calibration product be concentration be respectively 0GPLU/mL, 10GPLU/mL,
Anticardiolipin antibody IgG of 20GPLU/mL, 50GPLU/mL, 100GPLU/mL and 200GPLU/mL
Solution.
Concrete, anticardiolipin antibody IgG calibration product can use standard substance buffer by anticardiolipin antibody
IgG be configured to concentration be respectively 0GPLU/mL, 10GPLU/mL, 20GPLU/mL, 50GPLU/mL,
The solution of anticardiolipin antibody IgG of 100GPLU/mL and 200GPLU/mL.
This anticardiolipin antibody IgG chemiluminescence immune detection reagent kit is examined for anticardiolipin antibody IgG
During survey, utilize Full-automatic chemiluminescence immunoassay analysis meter that anticardiolipin antibody IgG calibration product are detected,
Draw standard curve, be built in computer software；Then test actual sample, calculate sample according to sample luminous value
This concentration；Finally anticardiolipin antibody IgG automatic chemiluminescence immunoassay system is carried out performance (spirit
Sensitivity, linear, precision, interference) evaluation.
This anticardiolipin antibody IgG chemiluminescence immune detection reagent kit can be exempted from Full-automatic chemiluminescence
Epidemic disease analyser is detection instrument, completes the detection this anticardiolipin antibody IgGization of anticardiolipin antibody IgG
Learning electrochemiluminescent immunoassay detection kit, through experiment, its detection sensitivity reaches 0.2GPLU/mL, relative to biography
The detection method sensitivity of anticardiolipin antibody IgG of system at least improves 10 times, this anticardiolipin antibody
The accuracy of detection of IgG chemiluminescence immune detection reagent kit is higher.
Additionally, this anticardiolipin antibody IgG chemiluminescence immune detection reagent kit also has the advantage that
1, selection acridinium ester is as marker material, and is applied to chemiluminescence immunoassay system, this luminous body
System is direct chemiluminescence, and compared with traditional enzyme-catalyzed chemical luminescence, this reaction need not the participation of enzyme, more
Add cost-effective；
2, select acridinium ester chemiluminescence immunoassay system range of linearity width, can reach 2GPLU/mL～
120GPLU/mL, and the inspection range of linearity of the detection method of traditional anticardiolipin antibody IgG is
10GPLU/mL～60GPLU/mL；
3, acridinium ester chemiluminescent immunoassay system repeatability is high, in batch and difference between batch is all within 5%,
This is that other chemiluminescence immunoassay system is unapproachable；
4, chemiluminescence immunoassay system has realized the quantitative of sample, soft to test by built-in standard curve
Part, only needs test sample just can directly obtain the concentration value of sample；
5, chemiluminescence immunoassay system can realize the interpolation of full-automation, reagent and sample instrument entirely
Complete, operate easier, decrease artificial error.
The preparation method of above-mentioned anticardiolipin antibody IgG chemiluminescence immune detection reagent kit as shown in Figure 1,
Comprise the steps:
Taking the suspension of carboxylated magnetic particle, Magneto separate is resuspended with MES buffer after removing supernatant, then adds
Enter EDC aqueous solution, the surface carboxyl groups of the magnetic particle of activated carboxyl, it is subsequently added into anticardiolipin antibody IgG
Monoclonal antibody, suspendible 2h～10h under room temperature, Magneto separate is resuspended with Tris buffer after removing supernatant, obtains
The coated carboxylated magnetic particle of anticardiolipin antibody IgG monoclonal antibody.
The concentration of MES (2-(N-morpholine) ethyl sulfonic acid) buffer be 0.02M, pH be 5.5.
The concentration of Tris buffer is 0.1M and is 8.0 containing 2%BSA, pH.
The concentration of EDC (1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimides) aqueous solution is
10mg/mL～20mg/mL, EDC are 0.05:0.1～1 with the ratio of carboxylated magnetic particle.
Preferably, in the coated carboxylated magnetic particle of anticardiolipin antibody IgG monoclonal antibody, anti-heart phosphorus
Fat IgG antibody monoclonal antibody is 1:25～35 with the ratio of carboxylated magnetic particle.
Preferably, the particle diameter of carboxylated magnetic particle is 0.05 μm～1 μm.
Take anticardiolipin antibody IgG monoclonal antibody, mix after adding carbonate buffer solution, being subsequently adding
Mixing after learning luminous marker, under room temperature, remove impurity after lucifuge reaction 1h～2h, obtains the anti-labelling of anti-human igg two
Chemiluminescent labels.
Order can overturn.
Carbonate buffer solution concentration be 0.1M, pH be 9.0～9.5,
The operation of remove impurity is centrifugal desalting column desalination, and concrete operations are: the most respectively by pure water and TBS buffering
Liquid (40mM Tris-HCl, 0.5%BSA, 1%NaCl, pH 8.0) processes centrifugal desalting column, finally adds
Enter the solution of the coated carboxylated magnetic particle of the anticardiolipin antibody IgG monoclonal antibody obtained, finally receive
Liquid in collection centrifuge tube.
Preferably, in the chemiluminescent labels of the anti-labelling of anti-human igg two, anticardiolipin antibody IgG Dan Ke
Grand antibody is 50:1～10 with the ratio of chemiluminescent labels.
Chemiluminescent labels can be luminol, different luminol, tris (bipyridine) ruthenium or acridinium ester.Wherein,
Chemiluminescent labels is preferably acridinium ester.
The coated carboxylated magnetic particle of anticardiolipin antibody IgG monoclonal antibody obtained and anti-human igg two
The chemiluminescent labels combination of anti-labelling i.e. can get the inspection of above-mentioned anticardiolipin antibody IgG chemiluminescence immunoassay
Test agent box.
This anticardiolipin antibody IgG chemiluminescence immune detection reagent kit is in use, in addition it is also necessary to chemistry is sent out
Light substrate solution and anticardiolipin antibody IgG calibration product.
Chemoluminescent substrate and anticardiolipin antibody IgG calibration product can be prepared voluntarily and obtain.
Chemoluminescent substrate includes A liquid and B liquid.A liquid can be H2O2Solution, B liquid can be NaOH
Solution.
In the present embodiment, A liquid be concentration be the H of 0.1mol/L2O2Solution, B liquid be concentration be 0.25mol/L
NaOH solution.
Concrete, anticardiolipin antibody IgG calibration product can use standard substance buffer by anticardiolipin antibody
IgG be configured to concentration be respectively 0GPLU/mL, 10GPLU/mL, 20GPLU/mL, 50GPLU/mL,
The solution of anticardiolipin antibody IgG of 100GPLU/mL and 200GPLU/mL.
The preparation method of this anticardiolipin antibody IgG chemiluminescence immune detection reagent kit is simple and convenient, system
The detection sensitivity of the anticardiolipin antibody IgG chemiluminescence immune detection reagent kit obtained is higher, has good
Application prospect.
It it is below specific embodiment.
Embodiment 1: the preparation of anticardiolipin antibody IgG chemiluminescence immune detection reagent kit
(1) preparation of the coated carboxylated magnetic particle of anticardiolipin antibody IgG monoclonal antibody:
Take containing the carboxylated magnetic particle (MagnaBind that 50mg particle diameter is 0.05 μm～1 μmTM, article No.
21353) suspension, Magneto separate removes supernatant, is that 5.5MES buffer is resuspended with 0.02M, pH, adds
The EDC aqueous solution of the 10mg/mL that 1mL is newly configured, activated magnetic beads surface carboxyl groups, add 4mg anti-heart phosphorus
Fat IgG antibody monoclonal antibody (biorbyt, article No. orb48780), suspendible 6h under room temperature, Magneto separate, goes
Except supernatant, it is resuspended to 1mg/mL with the Tris buffer that the 0.1M containing 2%BSA, pH are 8.0, obtains
The coated carboxylated magnetic particle of anticardiolipin antibody IgG monoclonal antibody, every bottle of 5mL subpackage is stored in 4
DEG C standby.
(2) preparation of the acridinium ester of the anti-labelling of anti-human igg two:
Take the anticardiolipin antibody IgG monoclonal antibody that 50 μ L concentration are 25mg/mL, add 150 μ L dense
Degree is the carbonate buffer solution that 0.1M, pH are 9.0～9.5, and mixing, being subsequently adding 1.5 μ L concentration is 5mg/mL
Acridinium ester solution mixing, under room temperature lucifuge reaction, after 1.5h take out, be centrifuged desalting column with the zeba of 2mL
Desalting processing, processes with pure water and TBS buffer the most respectively in desalination processes, is eventually adding
The acridinium ester solution of the anti-labelling of anti-human igg two arrived, the liquid collected in centrifuge tube is in control anti-human to preservation
The acridinium ester of the anti-labelling of IgG bis-, every bottle of 5mL subpackage be stored in 4 DEG C standby.
(3) preparation of anticardiolipin antibody IgG calibration product:
With standard substance buffer (40mM Tris-HCl, 0.5%BSA, 1%NaCl, pH 8.0) by the anti-heart
Phospholipid antibody IgG be configured to concentration be 0GPLU/mL, 10GPLU/mL, 20GPLU/mL,
50GPLU/mL, 100GPLU/mL and 200GPLU/mL, every bottle of 0.5mL subpackage lyophilizing, 4 DEG C of preservations
Standby.
Embodiment 2: anticardiolipin antibody IgG chemical luminous immune detection method
It is detection instrument with Full-automatic chemiluminescence immunoassay analysis meter (YHLO, article No. iFlash3000), side
Science of law pattern is that double antibody sandwich method, i.e. instrument are sequentially added into the sample of 50 μ L, the anticardiolipin of 50 μ L resists
The coated carboxylated magnetic particle of body IgG monoclonal antibody and the anticardiolipin antibody IgG process of 50 μ L
Liquid, after reaction 20min, then adds the coated acridinium ester of anticardiolipin antibody IgG of 50 μ L, reacts 20min
After, carrying out Magneto separate, reactant mixture is sent into darkroom by instrument, is sequentially added into luminous substrate A liquid (H2O2)
And B liquid (NaOH) carries out luminescence-producing reaction, finally record luminous value.
Embodiment 3: anticardiolipin antibody IgG chemiluminescence immune detection reagent kit performance evaluation
Use the method in embodiment 2 that anticardiolipin antibody IgG calibration product are detected, obtain drawing mark
Directrix curve is as shown in Figure 2.
Then to then testing actual sample, concentration of specimens is calculated according to sample luminous value.
The detection of sensitivity:
With reference to CLSI EP17-A file recommendation experimental program, calculate the chemiluminescence of anticardiolipin antibody IgG and exempt from
The sensitivity of epidemic disease detection kit, the sensitivity tried to achieve is 0.2GPLU/mL.
Linear detection:
It is 2GPLU/mL, 10GPLU/mL, 20GPLU/mL, 50GPLU/mL, 120GPLU/mL to concentration
Standard substance do linear analysis, calculate linearly dependent coefficient, and r=0.9996, it addition, this test kit is to anticardiolipin
The range of linearity of IgG antibody sample detection is 2GPLU/ml～120GPLU/mL.
Precision measures:
Taking concentration is 10GPLU/mL and two anticardiolipin antibody IgG samples of 100GPLU/mL, each
The each concentration of sample respectively do 3 parallel, detect with three batches of test kits, calculate test kit criticize interior and batch between
Difference, result shows that this test kit is criticized interior and difference between batch and is respectively less than 5%.
Interference is tested:
Take pooled serum to add chaff interference respectively and include: conjugated bilirubin, unconjugated bilirubin, hemoglobin,
Ascorbic acid, glyceride, adding proportion is carried out according to 1:20, measures pooled serum respectively and with the addition of each
After kind chaff interference, the measured value of pooled serum, calculates deviation therebetween, with ± 10% as tolerance interval.Knot
Fruit shows, interference all reaches the file standard of NCCLS, can be used for clinical laboratory's anticardiolipin antibody
The accurate evaluation of IgG situation.
Embodiment 4, the contrast experiment of anticardiolipin antibody IgG chemiluminescence immune detection reagent kit
It is 0.1GPLU/mL by chemical luminescence detection method and traditional enzyme linked immunosorbent assay to concentration respectively
And the anticardiolipin antibody IgG sample of 10GPLU/mL detects, two kinds of method detection sensitivities are compared,
Data are as shown in the table:
As can be seen from the above table, the sensitivity of chemical luminescence detection method relatively enzyme linked immunosorbent assay improves about
20 times.
Embodiment described above only have expressed the several embodiments of the present invention, and it describes more concrete and detailed,
But therefore can not be interpreted as the restriction to the scope of the claims of the present invention.It should be pointed out that, for this area
Those of ordinary skill for, without departing from the inventive concept of the premise, it is also possible to make some deformation and
Improving, these broadly fall into protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be with appended
Claim is as the criterion.
Claims (10)
1. an anticardiolipin antibody IgG chemiluminescence immune detection reagent kit, it is characterised in that including:
The coated carboxylated magnetic particle of anticardiolipin antibody IgG monoclonal antibody and the change of the anti-labelling of anti-human igg two
Learn luminous marker.
Anticardiolipin antibody IgG chemiluminescence immune detection reagent kit the most according to claim 1, its
It is characterised by, in the coated carboxylated magnetic particle of described anticardiolipin antibody IgG monoclonal antibody, described
Anticardiolipin antibody IgG monoclonal antibody is 1:25～35 with the ratio of described carboxylated magnetic particle.
Anticardiolipin antibody IgG chemiluminescence immune detection reagent kit the most according to claim 1, its
It is characterised by, in the chemiluminescent labels of the described anti-labelling of anti-human igg two, described anticardiolipin antibody IgG
Monoclonal antibody is 50:1～10 with the ratio of described chemiluminescent labels.
Anticardiolipin antibody IgG chemiluminescence immune detection reagent kit the most according to claim 1, its
Being characterised by, the particle diameter of described carboxylated magnetic particle is 0.05 μm～1 μm.
Anticardiolipin antibody IgG chemiluminescence immune detection reagent kit the most according to claim 1, its
Being characterised by, described chemiluminescent labels is luminol, different luminol, tris (bipyridine) ruthenium or acridinium ester.
Anticardiolipin antibody IgG chemiluminescence immune detection reagent kit the most according to claim 1, its
It is characterised by, also includes that Chemoluminescent substrate, described Chemoluminescent substrate include A liquid and B liquid.
Anticardiolipin antibody IgG chemiluminescence immune detection reagent kit the most according to claim 6, its
Being characterised by, described A liquid is H2O2Solution, described B liquid is NaOH solution.
Anticardiolipin antibody IgG chemiluminescence immune detection reagent kit the most according to claim 1, its
It is characterised by, also includes that anticardiolipin antibody IgG calibrates product.
Anticardiolipin antibody IgG chemiluminescence immune detection reagent kit the most according to claim 8, its
Be characterised by, described anticardiolipin antibody IgG calibration product be concentration be respectively 0GPLU/mL,
10GPLU/mL, 20GPLU/mL, 50GPLU/mL, 100GPLU/mL and 200GPLU/mL's is anti-
The solution of cardiolipin antibody IgG.
10. exempt from according to the anticardiolipin antibody IgG chemiluminescence according to any one of claim 1～9 for one kind
The preparation method of epidemic disease detection kit, it is characterised in that comprise the steps:
Taking the suspension of carboxylated magnetic particle, Magneto separate is resuspended with MES buffer after removing supernatant, then adds
Enter EDC aqueous solution, the surface carboxyl groups of the magnetic particle of activated carboxyl, it is subsequently added into anticardiolipin antibody IgG
Monoclonal antibody, suspendible 2h～10h under room temperature, Magneto separate is resuspended with Tris buffer after removing supernatant, obtains
The coated carboxylated magnetic particle of anticardiolipin antibody IgG monoclonal antibody；And
Take anticardiolipin antibody IgG monoclonal antibody, mix after adding carbonate buffer solution, being subsequently adding
Mixing after learning luminous marker, under room temperature, remove impurity after lucifuge reaction 1h～2h, obtains the anti-labelling of anti-human igg two
Chemiluminescent labels.
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| CN201610516757.5A CN105891194A (en) | 2016-06-30 | 2016-06-30 | Anticardiolipin antibody IgG chemiluminescence immunoassay kit and preparation method thereof |
| PCT/CN2017/080408 WO2018000903A1 (en) | 2016-06-30 | 2017-04-13 | Anti-cardiolipin antibody igg chemiluminescence immunoassay kit and method for preparing same |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018000903A1 (en) * | 2016-06-30 | 2018-01-04 | 深圳市亚辉龙生物科技股份有限公司 | Anti-cardiolipin antibody igg chemiluminescence immunoassay kit and method for preparing same |
| CN109342718A (en) * | 2018-09-29 | 2019-02-15 | 宁波奥丞生物科技有限公司 | A kind of magnetic microparticle chemiluminescence detection method |
| CN111007261A (en) * | 2019-12-23 | 2020-04-14 | 江苏三联生物工程有限公司 | Electrochemiluminescence kit for detecting RyR-Ab and preparation method thereof |
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| CN111007261A (en) * | 2019-12-23 | 2020-04-14 | 江苏三联生物工程有限公司 | Electrochemiluminescence kit for detecting RyR-Ab and preparation method thereof |
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