CN105954510A - Chemiluminescence immunodetection kit for anti-beta 2 glycoprotein I antibodies IgG and preparation method of kit - Google Patents
Chemiluminescence immunodetection kit for anti-beta 2 glycoprotein I antibodies IgG and preparation method of kit Download PDFInfo
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- CN105954510A CN105954510A CN201610510237.3A CN201610510237A CN105954510A CN 105954510 A CN105954510 A CN 105954510A CN 201610510237 A CN201610510237 A CN 201610510237A CN 105954510 A CN105954510 A CN 105954510A
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- G—PHYSICS
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- 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
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
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- G01N33/532—Production of labelled immunochemicals
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- 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
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
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Abstract
The invention discloses a chemiluminescence immunodetection kit for anti-beta 2 glycoprotein I antibodies IgG and a preparation method of the kit. The chemiluminescence immunodetection kit for the anti-beta 2 glycoprotein I antibodies IgG comprises beta 2 glycoprotein I antigen-coated chloracetylated magnetic particles and anti-human IgG secondary antibody-marked chemiluminescence labels. The anti-beta 2 glycoprotein I antibody IgG chemiluminescence immunodetection kit can complete detection of anti-beta 2 glycoprotein I antibodies IgG by taking a full-automatic chemiluminescence immunity analyzer as a detection tool. An experiment shows that the chemiluminescence immunodetection kit has the detection sensitivity of 2 AU/mL; compared with the conventional detection method for the anti-beta 2 glycoprotein I antibodies IgG, the chemiluminescence immunodetection kit disclosed by the invention has the advantage that the sensitivity is at least improved by 10 times. The chemiluminescence immunodetection kit for the anti-beta 2 glycoprotein I antibodies IgG is higher in detection precision.
Description
Technical field
The present invention relates to vitro detection field, particularly relate to a kind of anti-beta 2 glycoprotein I IgG antibody chemiluminescence
Immunity detection reagent and preparation method thereof.
Background technology
Beta 2 glycoprotein I (β 2-GPI), is also called human apolipoprotein H, confirms that in 1961 it is a kind of first
The plasma protein of 50kD.And determining its molecular biological characteristic in 1984, its function is as anti-heart phosphorus
The cofactor that fat antibody core phospholipid combines.
Antiphospholipid syndrome (APS) is to occur that in patient body high titre anti-phospholipid antibody (APL) is main
One non-organ specific autoimmune's property disease of feature, can individually fall ill, it is possible to other autoimmune
Disease occurs together, especially systemic lupus erythematosus (sle) (SLE).Its Major Clinical feature is repeatedly plasma viscosity
Bolt is formed, multiple organ ischemia and habitual abortion.Sydney in 2005 is held international thrombosis hemostasis association
(ISTH) meeting augments the anti-β2-GPI antibody positive first as one of APS laboratoary markers.
The main method of Clinical detection anti-beta 2 glycoprotein I IgG antibody is enzyme linked immunosorbent assay, but the method
There is also 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 anti-beta 2 glycoprotein I IgG antibody chemistry providing a kind of detection sensitivity higher is sent out
Light immunity detection reagent and preparation method thereof.
A kind of anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit, including: beta 2 glycoprotein I
Antigen coated carboxylated magnetic particle and the chemiluminescent labels of the anti-labelling of anti-human igg two.
In the carboxylated magnetic particle that described beta 2 glycoprotein I is antigen coated, described beta 2 glycoprotein I antigen with
The ratio of described carboxylated magnetic particle is 1:25～35.
In the chemiluminescent labels of the described anti-labelling of anti-human igg two, described anti-human igg two resists and describedization
The ratio learning luminous marker is 50:1～10.
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 anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit, also includes chemiluminescence
Substrate solution, described Chemoluminescent substrate includes exciting liquid and preexciting liquid.
Described preexciting liquid is H2O2Solution, described exciting liquid is NaOH solution.
Described anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit, also includes anti-β 2 sugar
Protein I IgG antibody calibration product.
Described anti-beta 2 glycoprotein I IgG antibody calibration product be concentration be respectively 0AU/mL, 10AU/mL,
The anti-beta 2 glycoprotein I IgG antibody of 20AU/mL, 50AU/mL, 100AU/mL and 200AU/mL molten
Liquid.
A kind of preparation method of above-mentioned anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit,
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 beta 2 glycoprotein I antigen,
Suspendible 2h～10h under room temperature, Magneto separate is resuspended with Tris buffer after removing supernatant, obtains beta 2 glycoprotein I
Antigen coated carboxylated magnetic particle；And
Take anti-human igg two to resist, mix, after being subsequently adding chemiluminescent labels after adding carbonate buffer solution
Mixing, under room temperature, remove impurity after lucifuge reaction 1h～2h, obtains the chemiluminescent labeling of the anti-labelling of anti-human igg two
Thing.
This anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit can be sent out with full-automatic chemical
Light immunity analysis instrument is detection instrument, completes this anti-beta2 Glycoprotein of detection of anti-beta 2 glycoprotein I IgG antibody
I IgG antibody chemiluminescence immune detection reagent kit, through experiment, its detection sensitivity reaches 2AU/mL,
At least improve 10 times relative to the detection method sensitivity of traditional anti-beta 2 glycoprotein I IgG antibody, this
The accuracy of detection of anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit is higher.
Accompanying drawing explanation
Fig. 1 is the system of the anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit of an embodiment
The flow chart of Preparation Method；
Wherein, tetragon reality frame is critical process, and hexagon frame is qualifying point.
Fig. 2 is the anti-beta 2 glycoprotein I IgG antibody 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 anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit of one embodiment, including: anti-
Human IgG two resists coated carboxylated magnetic particle and the chemiluminescent labels of the anti-labelling of anti-human igg two.
Preferably, in the carboxylated magnetic particle that beta 2 glycoprotein I is antigen coated, beta 2 glycoprotein I antigen with
The ratio of carboxylated magnetic particle is 1:25～35.
Preferably, in the chemiluminescent labels of the anti-labelling of anti-human igg two, anti-human igg two is anti-to be sent out with chemistry
The ratio of signal thing is 50:1～10.
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 anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit
Also include Chemoluminescent substrate.
Chemoluminescent substrate includes preexciting liquid and exciting liquid.Preexciting liquid can be H2O2Solution, excites
Liquid can be NaOH solution.
In the present embodiment, preexciting liquid be concentration be the H of 0.1mol/L2O2Solution, exciting liquid is that concentration is
The NaOH solution of 0.25mol/L.
In other examples, above-mentioned anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit
Also include that anti-beta 2 glycoprotein I IgG antibody calibrates product.
Anti-beta 2 glycoprotein I IgG antibody calibration product be concentration be respectively 0AU/mL, 10AU/mL, 20AU/mL,
The solution of the anti-beta 2 glycoprotein I IgG antibody of 50AU/mL, 100AU/mL and 200AU/mL.
Concrete, anti-beta 2 glycoprotein I IgG antibody calibration product can use standard substance buffer by anti-β 2 sugar
Protein I IgG antibody be configured to concentration be respectively 0AU/mL, 10AU/mL, 20AU/mL, 50AU/mL,
The solution of the anti-beta 2 glycoprotein I IgG antibody of 100AU/mL and 200AU/mL.
This anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit is used for anti-beta 2 glycoprotein I
During IgG antibody detection, utilize Full-automatic chemiluminescence immunoassay analysis meter antagonism beta 2 glycoprotein I IgG antibody fixed
Mark product detect, and draw standard curve, are built in computer software；Then actual sample is tested, according to sample
This luminous value calculates concentration of specimens；Finally antagonism beta 2 glycoprotein I IgG antibody Full-automatic chemiluminescence immunity divides
Analysis system carries out the evaluation of performance (sensitivity, linear, precision, interference).
This anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit can be sent out with full-automatic chemical
Light immunity analysis instrument is detection instrument, completes this anti-beta2 Glycoprotein of detection of anti-beta 2 glycoprotein I IgG antibody
I IgG antibody chemiluminescence immune detection reagent kit, through experiment, its detection sensitivity reaches 0.069
AU/mL, at least improves 10 relative to the detection method sensitivity of traditional anti-beta 2 glycoprotein I IgG antibody
Times, the accuracy of detection of this anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit is higher.
Additionally, this anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit also has following excellent
Point:
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 5AU/mL～
190AU/mL, and the inspection range of linearity of the detection method of traditional anti-beta 2 glycoprotein I IgG antibody is
0AU/mL～100AU/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 of above-mentioned anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit as shown in Figure 1
Method, comprises 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 beta 2 glycoprotein I antigen,
Suspendible 2h～10h under room temperature, Magneto separate is resuspended with Tris buffer after removing supernatant, obtains beta 2 glycoprotein I
Antigen coated carboxylated magnetic particle.
The concentration of MES (2-(N-morpholine) ethyl sulfonic acid) buffer be 0.02M, pH be 5.5.
The concentration of PBS 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 carboxylated magnetic particle that beta 2 glycoprotein I is antigen coated, beta 2 glycoprotein I antigen with
The ratio of carboxylated magnetic particle is 1:25～35.
Preferably, the particle diameter of carboxylated magnetic particle is 0.05 μm～1 μm.
Take anti-human igg two to resist, mix, after being subsequently adding chemiluminescent labels after adding carbonate buffer solution
Mixing, under room temperature, remove impurity after lucifuge reaction 1h～2h, obtains the chemiluminescent labeling of the anti-labelling of anti-human igg two
Thing.
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 antigen coated carboxylated magnetic particle of the beta 2 glycoprotein I obtained, finally collect in centrifuge tube
Liquid.
Preferably, in the chemiluminescent labels of the anti-labelling of anti-human igg two, anti-human igg two is anti-to be sent out with chemistry
The ratio of signal thing is 50:1～10.
Chemiluminescent labels can be luminol, different luminol, tris (bipyridine) ruthenium or acridinium ester.Wherein,
Chemiluminescent labels is preferably acridinium ester.
Carboxylated magnetic particle that the beta 2 glycoprotein I that obtains is antigen coated and the chemistry of the anti-labelling of anti-human igg two
Luminous marker combination i.e. can get above-mentioned anti-beta 2 glycoprotein I IgG antibody chemiluminescence immunoassay detectable
Box.
This anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit is in use, in addition it is also necessary to change
Learn luminous substrate liquid and anti-beta 2 glycoprotein I IgG antibody calibration product.
Chemoluminescent substrate and anti-beta 2 glycoprotein I IgG antibody calibration product can be prepared voluntarily and obtain.
Chemoluminescent substrate includes preexciting liquid and exciting liquid.Preexciting liquid can be H2O2Solution, excites
Liquid can be NaOH solution.
In the present embodiment, preexciting liquid be concentration be the H of 0.1mol/L2O2Solution, exciting liquid is that concentration is
The NaOH solution of 0.25mol/L.
Concrete, anti-beta 2 glycoprotein I IgG antibody calibration product can use standard substance buffer by anti-β 2 sugar
Protein I IgG antibody be configured to concentration be respectively 0AU/mL, 10AU/mL, 20AU/mL, 50AU/mL,
The solution of the anti-beta 2 glycoprotein I IgG antibody of 100AU/mL and 200AU/mL.
The preparation method of this anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit is the most square
Just, the detection sensitivity of the anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit prepared is higher,
Have a good application prospect.
It it is below specific embodiment.
Embodiment 1: the preparation of anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit
(1) preparation of the carboxylated magnetic particle that beta 2 glycoprotein I is antigen coated:
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 the anti-β of 4mg 2
Glycoprotein I antigen (Meridian, article No. A3C083H), suspendible 6h under room temperature, Magneto separate, remove supernatant,
It is resuspended to 1mg/mL with the Tris buffer that the 0.1M containing 2%BSA, pH are 8.0, obtains beta2 Glycoprotein
The coated carboxylated magnetic particle of I antigen, every bottle of 5mL subpackage be stored in 4 DEG C standby.
(2) preparation of the acridinium ester of the anti-labelling of anti-human igg two:
Taking the anti-human igg two that 50 μ L concentration are 25mg/mL to resist, adding 150 μ L concentration is 0.1M, pH
It is the carbonate buffer solution of 9.0～9.5, mixing, it is subsequently adding the acridinium ester that 1.5 μ L concentration are 5mg/mL molten
Liquid mixes, lucifuge reaction under room temperature, takes out, be centrifuged desalting column desalting processing with the zeba of 2mL after 1.5h,
Desalination processes processes with pure water and TBS buffer the most respectively, is eventually adding the anti-human igg obtained
The acridinium ester solution of two anti-labellings, the liquid collected in centrifuge tube is in control the anti-labelling of anti-human igg two to preservation
Acridinium ester, every bottle of 5mL subpackage be stored in 4 DEG C standby.
(3) preparation of anti-beta 2 glycoprotein I IgG antibody calibration product:
With standard substance buffer (40mM Tris-HCl, 0.5%BSA, 1%NaCl, pH 8.0) by anti-β 2
Glycoprotein I IgG antibody be configured to concentration be 0AU/mL, 10AU/mL, 20AU/mL, 50AU/mL,
100AU/mL and 200AU/mL, every bottle of 0.5mL subpackage ,-20 DEG C save backup.
Embodiment 2: anti-beta 2 glycoprotein I IgG antibody 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 5 μ L, the beta 2 glycoprotein I of 50 μ L
Antigen coated carboxylated magnetic particle and the anti-beta 2 glycoprotein I IgG antibody treatment fluid of 95 μ L, reaction
After 10min, then add the anti-coated acridinium ester of beta 2 glycoprotein I IgG antibody of 100 μ L, react 10min
After, carrying out Magneto separate, reactant mixture is sent into darkroom by instrument, is sequentially added into luminous substrate preexciting liquid
(H2O2) and exciting liquid (NaOH) carry out luminescence-producing reaction, finally record luminous value.
Embodiment 3: anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit performance evaluation
Use the antagonism beta 2 glycoprotein I IgG antibody calibration product of the method in embodiment 2 to detect, painted
Standard curve processed 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 anti-beta 2 glycoprotein I IgG antibody chemistry and send out
The sensitivity of light immunity detection reagent, the sensitivity tried to achieve is 0.069AU/mL.
Linear detection:
Be 5AU/mL to concentration, 42AU/mL, 79AU/mL, 116AU/mL, 153AU/mL,
190AU/mL standard substance do linear analysis, calculate linearly dependent coefficient, r=0.9991, it addition, this reagent
The range of linearity of box antagonism beta 2 glycoprotein I IgG antibody sample detection is 5AU/mL～190AU/mL.
Precision measures:
Taking concentration is 30AU/mL and 80AU/mL two anti-beta 2 glycoprotein I IgG antibody sample, 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 10%.
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 the anti-beta 2 glycoprotein I of clinical laboratory
The accurate evaluation of IgG antibody situation.
Embodiment 4, the contrast experiment of anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit
Respectively with chemical luminescence detection method and traditional enzyme linked immunosorbent assay to concentration be 0AU/mL, 15.3
AU/mL, anti-beta 2 glycoprotein I IgG antibody sample detect, two kinds of method detection sensitivities are compared, number
According to 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
30 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 anti-beta 2 glycoprotein I IgG antibody chemiluminescence immune detection reagent kit, it is characterised in that bag
Include: carboxylated magnetic particle that beta 2 glycoprotein I is antigen coated and the chemiluminescence mark of the anti-labelling of anti-human igg two
Note thing.
Anti-beta 2 glycoprotein I IgG antibody chemiluminescence immunoassay detectable the most according to claim 1
Box, it is characterised in that in the carboxylated magnetic particle that described beta 2 glycoprotein I is antigen coated, described β 2 sugar
Protein I antigen is 1:25～35 with the ratio of described carboxylated magnetic particle.
Anti-beta 2 glycoprotein I IgG antibody chemiluminescence immunoassay detectable the most according to claim 1
Box, it is characterised in that in the chemiluminescent labels of the described anti-labelling of anti-human igg two, described anti-human igg
Two anti-and described chemiluminescent labels ratios are 50:1～10.
Anti-beta 2 glycoprotein I IgG antibody chemiluminescence immunoassay detectable the most according to claim 1
Box, it is characterised in that the particle diameter of described carboxylated magnetic particle is 0.05 μm～1 μm.
Anti-beta 2 glycoprotein I IgG antibody chemiluminescence immunoassay detectable the most according to claim 1
Box, it is characterised in that described chemiluminescent labels is luminol, different luminol, tris (bipyridine) ruthenium or a word used for translation
Pyridine ester.
Anti-beta 2 glycoprotein I IgG antibody chemiluminescence immunoassay detectable the most according to claim 1
Box, it is characterised in that also include that Chemoluminescent substrate, described Chemoluminescent substrate include preexciting liquid
And exciting liquid.
Anti-beta 2 glycoprotein I IgG antibody chemiluminescence immunoassay detectable the most according to claim 6
Box, it is characterised in that described preexciting liquid is H2O2Solution, described exciting liquid is NaOH solution.
Anti-beta 2 glycoprotein I IgG antibody chemiluminescence immunoassay detectable the most according to claim 1
Box, it is characterised in that also include that anti-beta 2 glycoprotein I IgG antibody calibrates product.
Anti-beta 2 glycoprotein I IgG antibody chemiluminescence immunoassay detectable the most according to claim 8
Box, it is characterised in that described anti-beta 2 glycoprotein I IgG antibody calibration product be concentration be respectively 0AU/mL,
The anti-beta 2 glycoprotein I of 10AU/mL, 20AU/mL, 50AU/mL, 100AU/mL and 200AU/mL
The solution of IgG antibody.
10. send out according to the anti-beta 2 glycoprotein I IgG antibody chemistry according to any one of claim 1～9 for one kind
The preparation method of light immunity detection reagent, 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 beta 2 glycoprotein I antigen,
Suspendible 2h～10h under room temperature, Magneto separate is resuspended with Tris buffer after removing supernatant, obtains beta 2 glycoprotein I
Antigen coated carboxylated magnetic particle；And
Take anti-human igg two to resist, mix, after being subsequently adding chemiluminescent labels after adding carbonate buffer solution
Mixing, under room temperature, remove impurity after lucifuge reaction 1h～2h, obtains the chemiluminescent labeling of the anti-labelling of anti-human igg two
Thing.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610510237.3A CN105954510A (en) | 2016-06-30 | 2016-06-30 | Chemiluminescence immunodetection kit for anti-beta 2 glycoprotein I antibodies IgG and preparation method of kit |
| PCT/CN2017/080403 WO2018000899A1 (en) | 2016-06-30 | 2017-04-13 | Anti-β2 glycoprotein i antibody igg chemiluminescence immunoassay kit and preparation method thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610510237.3A CN105954510A (en) | 2016-06-30 | 2016-06-30 | Chemiluminescence immunodetection kit for anti-beta 2 glycoprotein I antibodies IgG and preparation method of kit |
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| WO (1) | WO2018000899A1 (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018000899A1 (en) * | 2016-06-30 | 2018-01-04 | 深圳市亚辉龙生物科技股份有限公司 | Anti-β2 glycoprotein i antibody igg chemiluminescence immunoassay kit and preparation method thereof |
| CN109100516A (en) * | 2018-09-19 | 2018-12-28 | 迪瑞医疗科技股份有限公司 | A kind of detection cytomegalovirus IgG antibody chemical luminescence immune assay determination reagent kit and preparation method thereof |
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| WO2018000899A1 (en) | 2018-01-04 |
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