CN106248943A - Antinuclear antibody chemiluminescence immune detection reagent kit and preparation method thereof - Google Patents
Antinuclear antibody chemiluminescence immune detection reagent kit and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of antinuclear antibody chemiluminescence immune detection reagent kit and preparation method thereof, antinuclear antibody chemiluminescence immune detection reagent kit includes: the coated carboxylated magnetic particle of antinuclear antibody monoclonal antibody and the chemiluminescent labels of inhibin labeling of monoclonal antibody.This antinuclear antibody chemiluminescence immune detection reagent kit can be with Full-automatic chemiluminescence immunoassay analysis meter for detection instrument, complete the detection this antinuclear antibody chemiluminescence immune detection reagent kit of antinuclear antibody, through experiment, its detection sensitivity reaches 4AU/mL, at least improve 10 times relative to the detection method sensitivity of traditional antinuclear antibody, the accuracy of detection of this antinuclear antibody chemiluminescence immune detection reagent kit is higher.
Description
Technical field
The present invention relates to vitro detection field, particularly relate to a kind of antinuclear antibody chemiluminescence immune detection reagent kit and
Preparation method.
Background technology
Antinuclear antibody (antinuclear antibody, ANA) also known as anti-antigen nucleic acid antibody, be one group by self eucaryon
Various composition deoxyribonucleoproteins (DNP), DNA, extractible nuclear antigen (ENA) and the RNA etc. of cell are as target antigen
The general name of autoantibody, be primarily present in serum.Antinuclear antibody in various autoimmune disease all in sun in various degree
Property rate, such as systemic lupus erythematosus (sle) (SLE, 95%～100%), rheumatoid arthritis (RA, 10%～20%), Combination
Connective tissue disease (MCTD, 80%～100%), dry syndrome (SS, 10%～40%), systemic scleroderma (85%～
90%), lupus hepatitis (95%～100%), primary biliary cirrhosis (95%～100%) etc..
The common methods of Clinical detection antinuclear antibody includes indirect immunofluorescence, enzyme linked immunosorbent assay, enzymatic chemistry
Luminescence method, but these methods all also exist deficiency, specific as follows shown.
One, indirect immunofluorescence
The ultimate principle of this method be combined with specific antibody antigen in microscope slide after form antigen antibody complex, continue
Be combined with antigen antibody complex by fluorescent antibody, form antigen-antibody fluorescent composition.Under fluorescence microscope, according to compound
The luminous situation of thing determines detected antibody.The method is evaluated: the fluorescence owing to being combined on antigen antibody complex resists
Body increases, and the fluorescent brightness sent is strong, thus its sensitivity is strong.But its deficiency is also apparent from:
(1) cannot be according to the non-specific identification of the size discrimination of molecular weight when analysis result；
(2) operate relative complex, need price fluorescence microscope costly, be difficult to promote at a lot of basic hospitals, the most less
It is applicable to the laboratory that specimen amount is more；
(3) background in fluoremetry is higher, and immunofluorence technic has certain difficulty for quantitative determination；
(4) result judges to need experienced professional, and the objectivity of analysis result is not enough；
(5) qualitative detection can only be carried out, it is impossible to quantitative determine.
Two, enzyme linked immunosorbent assay
Enzyme linked immunosorbent assay (ELISA) is widely used, 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 coated apparatus and reaction
Container, can only be divided into 12 batches, 6 batches, 8 batches or imposite first use in use, it is impossible to carry out independent, single part
Detection；
(2) reagent type used by quantitative determination is more, and each detectable will contain with reagent bottle, and often makes
Being required for changing imbibition nozzle during with a kind of reagent to be filled into respectively in the micropore of microwell plate, not only reagent bottle kind is many, filling
The operation of reagent is the most loaded down with trivial details；
(3) lack the corresponding mark to detection information, can only just will appreciate that by the mark checking test kit external packing box or know
Know product batch number and the effect duration information of detectable, and the information known is uncontrolled during detection, has the biggest
Randomness；
(4) detectable is in open space during detection, easily causes the cross-contamination between various reagent and shadow
Ring 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 the most loaded down with trivial details and multiple
Miscellaneous, 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 needing inspection
Survey 10 projects, then configuration and the use number of reagent must be 10 × 48/96 person-portions, if the most a sample needs to detect 10
Different project, it is also desirable to the reagent of configuration 10 × 48/96 person-portions, the shortcoming that there is inadequate economical rationality.
Three, chemoluminescence method
Chemoluminescence method can be divided into direct chemiluminescence and enzyme-catalyzed chemical luminescence by principle of luminosity.
Enzyme-catalyzed chemical luminescence mainly has horseradish peroxidase (HRP) and alkali phosphatase two kinds, but has certain office
Sex-limited, horseradish peroxidase major defect is: luminol, also can be by H2O2 in the case of not having horseradish peroxidase
Aoxidizing self luminous, background is of a relatively high, affects signal to noise ratio, and kinetics is complicated, and influence factor is many, and result is not sufficiently stable,
Obtain highly sensitive and plateau length substrate to be not easy.Alkali phosphatase major defect is: substrate reach plateau time
Between long, substrate cost is high, causes testing cost high, patient burden's weight.
Acridinium ester is compared enzyme-catalyzed chemical luminescence as the direct chemiluminescence of label and is had detailed advantage, mainly shows
: reaction need not catalyst, as long as alkaline environment can be carried out, is swift in response, and background luminescence is low, and signal to noise ratio is high, disturb because of
Element is few, and reagent stability is good, can be with two-point calibration, and system is simple, exciting liquid low cost, acridinium ester easily and protein bind, and
After connection, photon productivity does not reduces.
Summary of the invention
Based on this, it is necessary to provide antinuclear antibody chemiluminescence immune detection reagent kit that a kind of detection sensitivity is higher and
Its preparation method.
A kind of antinuclear antibody chemiluminescence immune detection reagent kit, including: the carboxylated magnetic that antinuclear antibody is antigen coated
Microgranule and the chemiluminescent labels of inhibin labeling of monoclonal antibody.
In one embodiment, in the carboxylated magnetic particle that described antinuclear antibody is antigen coated, described antinuclear antibody list
Clonal antibody is 1:20 ~ 100 with the ratio of described carboxylated magnetic particle.
In one embodiment, in the chemiluminescent labels of described inhibin labeling of monoclonal antibody, described anti-core resists
Body monoclonal antibody is 1:2 ~ 200 with the ratio of described chemiluminescent labels.
In one embodiment, the particle diameter of described carboxylated magnetic particle is 0.05 μm ~ 1 μm.
In one embodiment, described chemiluminescent labels is luminol, different luminol, tris (bipyridine) ruthenium or acridine
Ester.
In one embodiment, also include that Chemoluminescent substrate, described Chemoluminescent substrate include A liquid and B liquid.
In one embodiment, described A liquid is H2O2Solution, described B liquid is NaOH solution.
In one embodiment, also include that antinuclear antibody calibrates product.
In one embodiment, described antinuclear antibody calibration product be concentration be respectively 5AU/mL, 45AU/mL, 80AU/mL,
The solution of the antinuclear antibody of 160AU/mL, 320AU/mL and 640AU/mL.
The preparation method of a kind of above-mentioned antinuclear antibody chemiluminescence immune detection reagent kit, comprises the steps:
Taking the suspension of carboxylated magnetic particle, Magneto separate is resuspended with MES buffer after removing supernatant, is subsequently added into EDC aqueous solution,
The surface carboxyl groups of the magnetic particle of activated carboxyl, is subsequently added into antinuclear antibody antigen, suspendible 2h ~ 10h under room temperature, and Magneto separate is removed
Use Tris buffer resuspended after supernatant, obtain the carboxylated magnetic particle that antinuclear antibody is antigen coated；And
Take antinuclear antibody monoclonal antibody, mix after adding carbonate buffer solution, mix after being subsequently adding chemiluminescent labels,
Remove impurity after lucifuge reaction 1h ~ 2h, the chemiluminescent labels of the element labeling of monoclonal antibody that is inhibited under room temperature.
This antinuclear antibody chemiluminescence immune detection reagent kit can be with Full-automatic chemiluminescence immunoassay analysis meter for inspection
Survey instrument, completes the detection this antinuclear antibody chemiluminescence immune detection reagent kit of antinuclear antibody, and through experiment, it detects spirit
Sensitivity reaches 4AU/mL, at least improves 10 times relative to the detection method sensitivity of traditional antinuclear antibody, and this anti-core resists
The accuracy of detection of body chemiluminescence immune detection reagent kit is higher.
Accompanying drawing explanation
Fig. 1 is the flow chart of the preparation method of the antinuclear antibody chemiluminescence immune detection reagent kit of an embodiment；
Fig. 2 is the antinuclear 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, real with concrete below in conjunction with the accompanying drawings
Execute example the detailed description of the invention of the present invention is described in detail.Elaborate a lot of detail in the following description so that
Fully understand the present invention.But the present invention can implement to be much different from alternate manner described here, art technology
Personnel can do similar improvement in the case of intension of the present invention, and therefore the present invention is not embodied as by following public
Restriction.
The antinuclear antibody chemiluminescence immune detection reagent kit of one embodiment, including: the carboxylic that antinuclear antibody is antigen coated
The magnetic particle of base and the chemiluminescent labels of inhibin labeling of monoclonal antibody.
Preferably, in the carboxylated magnetic particle that antinuclear antibody is antigen coated, antinuclear antibody monoclonal antibody is with carboxylated
The ratio of magnetic particle be 1:20 ~ 100.
Preferably, in the chemiluminescent labels of inhibin labeling of monoclonal antibody, antinuclear antibody monoclonal antibody and change
The ratio learning luminous marker is 1:2 ~ 200.
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, chemiluminescence
Label is preferably acridinium ester.
In other examples, above-mentioned antinuclear antibody chemiluminescence immune detection reagent kit also includes chemical luminous substrate
Liquid.
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 that the NaOH of 0.25mol/L is molten
Liquid.
In other examples, above-mentioned antinuclear antibody chemiluminescence immune detection reagent kit also includes that antinuclear antibody is calibrated
Product.
Antinuclear antibody calibration product be concentration be respectively 5AU/mL, 45AU/mL, 80AU/mL, 160AU/mL, 320AU/mL and
The solution of the antinuclear antibody of 640AU/mL.
Concrete, antinuclear antibody calibration product can use standard substance buffer that antinuclear antibody is configured to concentration and be respectively
The solution of the antinuclear antibody of 5AU/mL, 45AU/mL, 80AU/mL, 160AU/mL, 320AU/mL and 640AU/mL.
This antinuclear antibody chemiluminescence immune detection reagent kit, when antinuclear antibody detects, utilizes full-automatic chemical to send out
Antinuclear antibody calibration product are detected by light immunity analysis instrument, draw standard curve, are built in computer software；Then reality is tested
Sample, calculates concentration of specimens according to sample luminous value；Finally antinuclear antibody automatic chemiluminescence immunoassay system is carried out
The evaluation of performance (sensitivity, linear, precision, interference).
This antinuclear antibody chemiluminescence immune detection reagent kit can be with Full-automatic chemiluminescence immunoassay analysis meter for inspection
Survey instrument, completes the detection this antinuclear antibody chemiluminescence immune detection reagent kit of antinuclear antibody, and through experiment, it detects spirit
Sensitivity reaches 4AU/mL, at least improves 10 times relative to the detection method sensitivity of traditional antinuclear antibody, and this anti-core resists
The accuracy of detection of body chemiluminescence immune detection reagent kit is higher.
Additionally, this antinuclear antibody 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, and this luminescence system is directization
Learning luminescence, compared with traditional enzyme-catalyzed chemical luminescence, this reaction need not the participation of enzyme, more cost-effective；
2, select the chemiluminescence immunoassay system range of linearity width of acridinium ester, 4AU/mL ~ 500AU/mL can be reached, and pass
The inspection range of linearity of the detection method of the antinuclear antibody of system is 20pg/mL ~ 1000pg/mL；
3, acridinium ester chemiluminescent immunoassay system repeatability is high, and in batch and difference between batch is all within 5%, this is other chemistry
Luminescence immunoassay system is unapproachable；
4, chemiluminescence immunoassay system has realized the quantitative of sample, by built-in standard curve to test software, only needs to survey
Sample originally just can directly obtain the concentration value of sample；
5, chemiluminescence immunoassay system can realize the interpolation of full-automation, reagent and sample has instrument to complete entirely, operation
Easier, decrease artificial error.
The preparation method of above-mentioned antinuclear antibody chemiluminescence immune detection reagent kit as shown in Figure 1, comprises the steps:
Taking the suspension of carboxylated magnetic particle, Magneto separate is resuspended with MES buffer after removing supernatant, is subsequently added into EDC aqueous solution,
The surface carboxyl groups of the magnetic particle of activated carboxyl, is subsequently added into antinuclear antibody antigen, suspendible 2h ~ 10h under room temperature, and Magneto separate is removed
Use Tris buffer resuspended after supernatant, obtain the carboxylated magnetic particle of antinuclear antibody antigen quilt.
MES(2-(N-morpholine) ethyl sulfonic acid) concentration of buffer be 0.02M, pH be 5.5.
The concentration of Tris buffer is 0.1M and is 8.0 containing 2%BSA, pH.
EDC(1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimides) concentration of 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 antinuclear antibody is antigen coated, antinuclear antibody antigen is micro-with carboxylated magnetic
The ratio of grain is 1:20 ~ 100.
Preferably, the particle diameter of carboxylated magnetic particle is 0.05 μm ~ 1 μm.
Take antinuclear antibody monoclonal antibody, mix, after being subsequently adding chemiluminescent labels after adding carbonate buffer solution
Mixing, remove impurity after lucifuge reaction 1h ~ 2h, the chemiluminescent labels of the element labeling of monoclonal antibody that is inhibited under room temperature.
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 with pure water and TBS buffer (40 mM
Tris-HCl, 0.5% BSA, 1% NaCl, pH 8.0) process centrifugal desalting column, it is eventually adding the antinuclear antibody monoclonal obtained
The solution of the coated carboxylated magnetic particle of antibody, finally collects the liquid in centrifuge tube.
Preferably, in the chemiluminescent labels of inhibin labeling of monoclonal antibody, antinuclear antibody monoclonal antibody and change
The ratio learning luminous marker is 1:2 ~ 200.
Chemiluminescent labels can be luminol, different luminol, tris (bipyridine) ruthenium or acridinium ester.Wherein, chemiluminescence
Label is preferably acridinium ester.
The coated carboxylated magnetic particle of antinuclear antibody monoclonal antibody that obtains and inhibin labeling of monoclonal antibody
Chemiluminescent labels combination i.e. can get above-mentioned antinuclear antibody chemiluminescence immune detection reagent kit.
This antinuclear antibody chemiluminescence immune detection reagent kit is in use, in addition it is also necessary to Chemoluminescent substrate and anti-core
Antibody calibration product.
Chemoluminescent substrate and antinuclear antibody 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 that the NaOH of 0.25mol/L is molten
Liquid.
Concrete, antinuclear antibody calibration product can use standard substance buffer that antinuclear antibody is configured to concentration and be respectively
The solution of the antinuclear antibody of 5AU/mL, 45AU/mL, 80AU/mL, 160AU/mL, 320AU/mL and 640AU/mL.
The preparation method of this antinuclear antibody chemiluminescence immune detection reagent kit is simple and convenient, the antinuclear antibody prepared
The detection sensitivity learning electrochemiluminescent immunoassay detection kit is higher, has a good application prospect.
It it is below specific embodiment.
Embodiment 1: the preparation of antinuclear antibody chemiluminescence immune detection reagent kit
(1) preparation of the coated carboxylated magnetic particle of antinuclear antibody monoclonal antibody:
Take containing carboxylated magnetic particle (MagnaBind, the article No. 21353) suspension that 50mg particle diameter is 0.05 μm ~ 1 μm,
Magneto separate removes supernatant, uses 0.02 M, and pH is that 5.5 MES buffer are resuspended, and the EDC adding 10mg/mL newly configured for 1mL is water-soluble
Liquid, activated magnetic beads surface carboxyl groups, add 4mg antinuclear antibody antigen, suspendible 6h under room temperature, Magneto separate, remove supernatant, with containing 2%
The 0.1M of BSA, pH be 8.0 Tris buffer be resuspended to 1mg/mL, obtain the carboxylated magnetic particle of antinuclear antibody antigen quilt,
Every bottle of 5mL subpackage be stored in 4 DEG C standby.
(2) preparation of the acridinium ester of inhibin labeling of monoclonal antibody:
Take the antinuclear antibody monoclonal antibody that 50 μ L concentration are 25mg/mL, add 150 μ L concentration be 0.1M, pH be 9.0 ~ 9.5
Carbonate buffer solution, mixing, it is subsequently adding the acridinium ester solution mixing that 1.5 μ L concentration are 5mg/mL, under room temperature, lucifuge is reacted,
Take out after 1.5h, be centrifuged desalting column desalting processing with the zeba of 2mL, the most respectively by pure water and TBS buffering in desalination processes
Liquid processes, and is eventually adding the acridinium ester solution of the inhibin labeling of monoclonal antibody obtained, and collects the liquid in centrifuge tube
Be in control the acridinium ester of inhibin labeling of monoclonal antibody to preserving, every bottle of 5mL subpackage be stored in 4 DEG C standby.
(3) preparation of antinuclear antibody calibration product:
Antinuclear antibody is configured to dense with standard substance buffer (40 mM Tris-HCl, 0.5% BSA, 1% NaCl, pH 8.0)
Degree is 5AU/mL, 45AU/mL, 80AU/mL, 160AU/mL, 320AU/mL and 640AU/mL, every bottle of 0.5 mL subpackage lyophilizing, 4
DEG C save backup.
Embodiment 2: antinuclear antibody chemical luminous immune detection method
Being detection instrument with Full-automatic chemiluminescence immunoassay analysis meter (YHLO, article No. iFlash3000), methodology pattern is double
Antibody sandwich, i.e. instrument be sequentially added into the antinuclear antibody antigen quilt of the sample of 5 μ L, 50 μ L carboxylated magnetic particle and
The antinuclear antibody treatment fluid of 50 μ L, after reaction 10min, then adds the coated acridinium ester of antinuclear antibody of 100 μ L, reacts 10min
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) enter
Row luminescence-producing reaction, finally records luminous value.
Embodiment 3: antinuclear antibody chemiluminescence immune detection reagent kit performance evaluation
Use the method in embodiment 2 that antinuclear antibody calibration product are detected, obtain drawing standard curve 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 sensitive of antinuclear antibody chemiluminescence immune detection reagent kit
Degree, the sensitivity tried to achieve is 4AU/ mL.
Linear detection:
Be 10AU/mL to concentration, 60AU/mL, 102AU/mL, 190AUpg/mL, 380AU/mL standard substance do linear point
Analysis, calculates linearly dependent coefficient, and r=0.9922, it addition, this test kit is 0AU/ to the range of linearity of antinuclear antibody sample detection
ml ~400AU/mL。
Precision measures:
Taking concentration is 160AU/mL and two antinuclear antibody samples of 342AU/mL, each concentration of each sample respectively do 3 parallel, use
Three batches of test kits detect, and calculate test kit and criticize interior and difference between batch, and 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, glycerol
Ester, adding proportion is carried out according to 1:20, the measured value of pooled serum after measuring pooled serum respectively and with the addition of various chaff interference,
Calculate deviation therebetween, with ± 10% as tolerance interval.Result shows, interference all reaches the files-designated of NCCLS
Standard, can be used for the accurate evaluation of clinical laboratory's antinuclear antibody situation.
Embodiment 4, the contrast experiment of antinuclear antibody chemiluminescence immune detection reagent kit
Be 10AU/mL by chemical luminescence detection method and traditional enzyme linked immunosorbent assay to concentration respectively, 60AU/mL,
The antinuclear antibody sample of 102AU/mL, 190AUpg/mL, 380AU/mL detects, and 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 20 times.
Embodiment described above only have expressed the several embodiments of the present invention, and it describes more concrete and detailed, but also
Therefore the restriction to the scope of the claims of the present invention can not be interpreted as.It should be pointed out that, for those of ordinary skill in the art
For, without departing from the inventive concept of the premise, it is also possible to make some deformation and improvement, these broadly fall into the guarantor of the present invention
Protect scope.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.
Claims (10)
1. an antinuclear antibody chemiluminescence immune detection reagent kit, it is characterised in that including: the carboxylic that antinuclear antibody is antigen coated
The magnetic particle of base and the chemiluminescent labels of inhibin labeling of monoclonal antibody.
Antinuclear antibody chemiluminescence immune detection reagent kit the most according to claim 1, it is characterised in that described anti-core resists
In the coated carboxylated magnetic particle of isoantigen, described antinuclear antibody antigen is 1:20 with the ratio of described carboxylated magnetic particle
~100。
Antinuclear antibody chemiluminescence immune detection reagent kit the most according to claim 1, it is characterised in that described inhibin
In the chemiluminescent labels of labeling of monoclonal antibody, described antinuclear antibody monoclonal antibody and described chemiluminescent labels
Ratio is 1:2 ~ 200.
Antinuclear antibody chemiluminescence immune detection reagent kit the most according to claim 1, it is characterised in that described carboxylated
The particle diameter of magnetic particle be 0.05 μm ~ 1 μm.
Antinuclear antibody chemiluminescence immune detection reagent kit the most according to claim 1, it is characterised in that described chemistry is sent out
Signal thing is luminol, different luminol, tris (bipyridine) ruthenium or acridinium ester.
Antinuclear antibody chemiluminescence immune detection reagent kit the most according to claim 1, it is characterised in that also include chemistry
Luminous substrate liquid, described Chemoluminescent substrate includes A liquid and B liquid.
Antinuclear antibody chemiluminescence immune detection reagent kit the most according to claim 6, it is characterised in that described A liquid is
H2O2Solution, described B liquid is NaOH solution.
Antinuclear antibody chemiluminescence immune detection reagent kit the most according to claim 1, it is characterised in that also include anti-core
Antibody calibration product.
Antinuclear antibody chemiluminescence immune detection reagent kit the most according to claim 8, it is characterised in that described anti-core resists
Body calibration product are the anti-core that concentration is respectively 5AU/mL, 45AU/mL, 80AU/mL, 160AU/mL, 320AU/mL and 640AU/mL
The solution of antibody.
10. the preparation according to the antinuclear antibody chemiluminescence immune detection reagent kit according to any one of claim 1 ~ 9
Method, 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, is subsequently added into EDC aqueous solution,
The surface carboxyl groups of the magnetic particle of activated carboxyl, is subsequently added into antinuclear antibody antigen, suspendible 2h ~ 10h under room temperature, and Magneto separate is removed
Use Tris buffer resuspended after supernatant, obtain the carboxylated magnetic particle that antinuclear antibody is antigen coated；And take antinuclear antibody list
Clonal antibody, mixes after adding carbonate buffer solution, mixes after being subsequently adding chemiluminescent labels, lucifuge reaction 1h under room temperature
Remove impurity after ~ 2h, the chemiluminescent labels of the element labeling of monoclonal antibody that is inhibited.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108333347A (en) * | 2017-01-19 | 2018-07-27 | 深圳市新产业生物医学工程股份有限公司 | Antinuclear antibodies target antigen conjugate reagent, preparation method, the kit comprising it and application |
| CN115656154A (en) * | 2022-10-26 | 2023-01-31 | 广东优尼德生物科技有限公司 | anti-RNP antibody chemiluminescence detection kit based on recombinant RNP multiple antigens |
| CN115656153A (en) * | 2022-10-26 | 2023-01-31 | 广东优尼德生物科技有限公司 | Antinuclear antibody spectrum detection kit based on acridinium ester chemiluminescence |
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