CN106053836A - Endometrial antibody chemiluminescence immunoassay kit and preparation method thereof - Google Patents
Endometrial antibody chemiluminescence immunoassay kit and preparation method thereof Download PDFInfo
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- CN106053836A CN106053836A CN201610503834.3A CN201610503834A CN106053836A CN 106053836 A CN106053836 A CN 106053836A CN 201610503834 A CN201610503834 A CN 201610503834A CN 106053836 A CN106053836 A CN 106053836A
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- 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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6854—Immunoglobulins
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- 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
- G01N33/543—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
- G01N33/54326—Magnetic particles
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- G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/36—Gynecology or obstetrics
- G01N2800/367—Infertility, e.g. sperm disorder, ovulatory dysfunction
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Abstract
The invention discloses an endometrial antibody chemiluminescence immunoassay kit and a preparation method thereof. The endometrial antibody chemiluminescence immunoassay kit comprises carboxylated magnetic particles enveloped by endometrial antibody recombinant protein, and a chemiluminescence marker marked by anti-human immunoglobulin. The endometrial antibody chemiluminescence immunoassay kit can complete the assay of an endometrial antibody by taking a full-automatic chemiluminescence immunity analyzer as an assay tool. Through experiments, the assay sensitivity of the endometrial antibody chemiluminescence immunoassay kit can reach 1U/L, and can be improved by at least ten times compared with that of the traditional endometrial antibody assay method, and the assay precision of the endometrial antibody chemiluminescence immunoassay kit is higher.
Description
Technical field
The present invention relates to vitro detection field, particularly relate to a kind of anti-endometrial antibodies chemiluminescence immunoassay detectable
Box and preparation method thereof.
Background technology
Anti-endometrial antibodies (AEAb) is a kind of autoantibody, and targeted antigen is in endometrial glandular epithelial cells
A kind of progestogen dependent protein, entopic endometrium is to body no antigen, but the endometrium of dystopy can conduct
Antigenic stimulus body immune system, produces special anti-endometrial antibodies not only anti-with ectopic endometrium generation antigen-antibody
Should, also can combining with the antigen site in entopic uterine veil cell simultaneously, activating complement system, local produces immunity
Pathological change, directly affects the function of endometrial gland, thus causes implantation of ovum failed or sterile, finally with inapparent
Early abortion and come to an end.Therefore anti-endometrial antibodies detection also has important meaning to clinical diagnosis with treatment sterility and infertility patient
Justice.
The common methods of Clinical detection anti-endometrial antibodies has enzyme linked immunosorbent assay, enzyme-catalyzed chemical luminescence method at present,
But in place of these methods all also exist some shortcomings.
One, 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 with anti-
Answer container, 12 batches, 6 batches, 8 batches or imposite first use can only be divided in use, it is impossible to carry out independent, single
The detection of person-portion；
(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
Individual different project, it is also desirable to the reagent of configuration 10 × 48/96 person-portions, the shortcoming that there is inadequate economical rationality.
Two, 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 the anti-endometrial antibodies chemiluminescence immunoassay detection examination that a kind of detection sensitivity is higher
Agent box and preparation method thereof.
A kind of anti-endometrial antibodies chemiluminescence immune detection reagent kit, including: anti-endometrial antibodies recombiant protein
Coated carboxylated magnetic particle and the chemiluminescent labels of human immunoglobulins's labelling.
In one embodiment, in the coated carboxylated magnetic particle of described anti-endometrial antibodies recombiant protein, described
Anti-endometrial antibodies recombiant protein is 1:25 ~ 35 with the ratio of described carboxylated magnetic particle.
In one embodiment, in the chemiluminescent labels of described human immunoglobulins's labelling, described anti-human immunity
Globulin is 50:1 ~ 10 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 anti-endometrial antibodies calibrates product.
In one embodiment, described anti-endometrial antibodies calibration product be concentration be respectively 1U/L, 10U/L, 100U/L,
The solution of the anti-endometrial antibodies of 500U/L, 1000U/L and 2000U/L.
The preparation method of a kind of above-mentioned anti-endometrial antibodies chemiluminescence immune detection reagent kit, including walking as follows
Rapid:
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 anti-endometrial antibodies recombiant protein, suspendible 2h ~ 10h under room temperature,
Magneto separate is resuspended with Tris buffer after removing supernatant, obtains the coated carboxylated magnetic of anti-endometrial antibodies recombiant protein micro-
Grain；And
Take human immunoglobulins, mix after adding carbonate buffer solution, mix after being subsequently adding chemiluminescent labels, room temperature
Remove impurity after lower lucifuge reaction 1h ~ 2h, obtains the chemiluminescent labels of human immunoglobulins's labelling.
This anti-endometrial antibodies chemiluminescence immune detection reagent kit can be with automatic chemiluminescence immunoassay
Instrument is detection instrument, completes the detection this anti-endometrial antibodies chemiluminescence immunoassay detectable of anti-endometrial antibodies
Box, through experiment, its detection sensitivity reaches 1U/L, relative to traditional anti-endometrial antibodies detection method sensitivity extremely
Improve 10 times less, the accuracy of detection of this anti-endometrial antibodies chemiluminescence immune detection reagent kit is higher.
Accompanying drawing explanation
Fig. 1 is the flow process of the preparation method of the anti-endometrial antibodies chemiluminescence immune detection reagent kit of an embodiment
Figure；
Fig. 2 is the anti-endometrial antibodies 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 anti-endometrial antibodies chemiluminescence immune detection reagent kit of one embodiment, including: anti-endometrial antibodies
The coated carboxylated magnetic particle of recombiant protein and the chemiluminescent labels of human immunoglobulins's labelling.
Preferably, in the coated carboxylated magnetic particle of anti-endometrial antibodies recombiant protein, anti-endometrial antibodies weight
Histone is 1:25 ~ 35 with the ratio of carboxylated magnetic particle.
Preferably, in the chemiluminescent labels of human immunoglobulins's labelling, human immunoglobulins and chemiluminescence
The ratio of label 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, chemiluminescence
Label is preferably acridinium ester.
In other examples, above-mentioned anti-endometrial antibodies chemiluminescence immune detection reagent kit also includes that chemistry is sent out
Light substrate solution.
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 anti-endometrial antibodies chemiluminescence immune detection reagent kit also includes anti-uterus
Interior membrane antibody calibration product.
Anti-endometrial antibodies calibration product be concentration be respectively 1U/L, 10U/L, 100U/L, 500U/L, 1000U/L and
The solution of the anti-endometrial antibodies of 2000U/L.
Concrete, anti-endometrial antibodies calibration product can use standard substance buffer anti-endometrial antibodies to be configured to
Concentration is respectively the solution of the anti-endometrial antibodies of 1U/L, 10U/L, 100U/L, 500U/L, 1000U/L and 2000U/L.
This anti-endometrial antibodies chemiluminescence immune detection reagent kit, when anti-endometrial antibodies detects, utilizes
Anti-endometrial antibodies calibration product are detected by Full-automatic chemiluminescence immunoassay analysis meter, draw standard curve, are built in electricity
Brain software；Then test actual sample, calculate concentration of specimens according to sample luminous value；Finally full-automatic to anti-endometrial antibodies
Chemiluminescence immunoassay system carries out the evaluation of performance (sensitivity, linear, precision, interference).
This anti-endometrial antibodies chemiluminescence immune detection reagent kit can be with automatic chemiluminescence immunoassay
Instrument is detection instrument, completes the detection this anti-endometrial antibodies chemiluminescence immunoassay detectable of anti-endometrial antibodies
Box, through experiment, its detection sensitivity reaches 1U/L, relative to traditional anti-endometrial antibodies detection method sensitivity extremely
Improve 10 times less, the accuracy of detection of this anti-endometrial antibodies chemiluminescence immune detection reagent kit is higher.
Additionally, this anti-endometrial antibodies 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, 1U/L ~ 1000U/L can be reached, and traditional
The inspection range of linearity of the detection method of anti-endometrial antibodies is 20U/L ~ 1000U/L；
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 anti-endometrial antibodies chemiluminescence immune detection reagent kit as shown in Figure 1, including such as
Lower step:
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 anti-endometrial antibodies recombiant protein, suspendible 2h ~ 10h under room temperature,
Magneto separate is resuspended with Tris buffer after removing supernatant, obtains the coated carboxylated magnetic of anti-endometrial antibodies recombiant protein micro-
Grain.
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 coated carboxylated magnetic particle of anti-endometrial antibodies recombiant protein, anti-endometrial antibodies weight
Histone 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 human immunoglobulins, mix after adding carbonate buffer solution, mix after being subsequently adding chemiluminescent labels,
Under room temperature, remove impurity after lucifuge reaction 1h ~ 2h, obtains the chemiluminescent labels of human immunoglobulins's labelling.
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 anti-endometrial antibodies obtained
The solution of the coated carboxylated magnetic particle of recombiant protein, finally collects the liquid in centrifuge tube.
Preferably, in the chemiluminescent labels of human immunoglobulins's labelling, anti-endometrial antibodies recombiant protein with
The ratio of chemiluminescent labels is 50:1 ~ 10.
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 anti-endometrial antibodies recombiant protein obtained and human immunoglobulins's labelling
Chemiluminescent labels combination i.e. can get above-mentioned anti-endometrial antibodies chemiluminescence immune detection reagent kit.
This anti-endometrial antibodies chemiluminescence immune detection reagent kit is in use, in addition it is also necessary to Chemoluminescent substrate
Product are calibrated with anti-endometrial antibodies.
Chemoluminescent substrate and anti-endometrial antibodies 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, anti-endometrial antibodies calibration product can use standard substance buffer anti-endometrial antibodies to be configured to
Concentration is respectively the solution of the anti-endometrial antibodies of 1U/L, 10U/L, 100U/L, 500U/L, 1000U/L and 2000U/L.
The preparation method of this anti-endometrial antibodies chemiluminescence immune detection reagent kit is simple and convenient, the anti-son prepared
The detection sensitivity of Endometrium antibody chemical luminescence immunity detection reagent is higher, has a good application prospect.
It it is below specific embodiment.
Embodiment 1: the preparation of anti-endometrial antibodies chemiluminescence immune detection reagent kit
(1) preparation of the coated carboxylated magnetic particle of anti-endometrial antibodies recombiant protein:
Taking containing carboxylated magnetic particle (MagnaBind21353) suspension that 50mg particle diameter is 0.05 μm ~ 1 μm, Magneto separate goes
Supernatant, uses 0.02 M, and pH is that 5.5 MES buffer are resuspended, adds the EDC aqueous solution of 10mg/mL newly configured for 1mL, activates magnetic
Bead surface carboxyl, addition 4mg anti-endometrial antibodies recombiant protein (biorbyt, article No. orb48780), suspendible 6h under room temperature,
Magneto separate, removes supernatant, is resuspended to 1mg/mL with the Tris buffer that the 0.1M containing 2%BSA, pH are 8.0, obtains anti-intrauterine
The coated carboxylated magnetic particle of membrane antibody recombiant protein, every bottle of 5mL subpackage be stored in 4 DEG C standby.
(2) preparation of the acridinium ester of human immunoglobulins's labelling:
Taking 50 μ L concentration is the human immunoglobulins of 25mg/mL, add 150 μ L concentration be 0.1M, pH be the carbonic acid of 9.0 ~ 9.5
Salt buffer, mixing, it is subsequently adding the acridinium ester solution mixing that 1.5 μ L concentration are 5mg/mL, lucifuge reaction under room temperature, after 1.5h
Take out, be centrifuged desalting column desalting processing with the zeba of 2mL, desalination processes is carried out with pure water and TBS buffer the most respectively
Process, be eventually adding the acridinium ester solution of the human immunoglobulins's labelling obtained, collect the liquid in centrifuge tube to preserving pipe
Obtain the acridinium ester of human immunoglobulins's labelling, every bottle of 5mL subpackage be stored in 4 DEG C standby.
(3) preparation of anti-endometrial antibodies calibration product:
With standard substance buffer (40 mM Tris-HCl, 0.5% BSA, 1% NaCl, pH 8.0), anti-endometrial antibodies is joined
Being set to concentration is 0U/L, 10U/L, 100U/L, 500U/L, 1000U/L and 2000U/L, every bottle of 0.5 mL subpackage lyophilizing, 4 DEG C of guarantors
Deposit standby.
Embodiment 2: anti-endometrial antibodies chemical luminous immune detection method
It is detection instrument with Full-automatic chemiluminescence immunoassay analysis meter (YHLO, article No. iFlash3000), between methodology pattern is
Connect immunization, i.e. instrument be sequentially added into the sample of 50 μ L, 50 μ L anti-endometrial antibodies recombiant protein coated carboxylated
Magnetic particle and the anti-endometrial antibodies treatment fluid of 50 μ L, after reacting 20 min, then add the anti-human immune globulin of 50 μ L
White acridinium ester, after reacting 20 min, carries out Magneto separate, and reactant mixture is sent into darkroom by instrument, is sequentially added into luminous substrate A liquid
(H2O2) and B liquid (NaOH) carry out luminescence-producing reaction, finally record luminous value.
Embodiment 3: anti-endometrial antibodies chemiluminescence immune detection reagent kit performance evaluation
Use the method in embodiment 2 that anti-endometrial antibodies calibration product are detected, obtain drawing standard curve such as Fig. 2 institute
Show.
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-endometrial antibodies chemiluminescence immune detection reagent kit
Sensitivity, the sensitivity tried to achieve is 1U/L.
Linear detection:
It is that 1U/L, 10U/L, 100U/L, 500U/L, 1000U/L and 2000U/L standard substance do linear analysis to concentration, calculates line
Property correlation coefficient, r=0.9996, it addition, this test kit to the range of linearity of anti-endometrial antibodies sample detection be 1U/L ~
1000U/L。
Precision measures:
Taking concentration is 50U/L and two anti-endometrial antibodies samples of 500U/L, each concentration of each sample respectively do 3 parallel,
Detecting with three batches of test kits, calculate test kit and criticize interior and difference between batch, result shows that this test kit is criticized interior and difference between batch is the least
In 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 anti-endometrial antibodies situation.
Embodiment 4, the contrast experiment of anti-endometrial antibodies chemiluminescence immune detection reagent kit
Be 0 by chemical luminescence detection method and traditional enzyme linked immunosorbent assay to concentration respectively, the anti-endometrium of 50U/L resists
Body sample detects, and two kinds of method detection sensitivities are compared, and 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 more than 10 times.
Embodiment described above only have expressed the several embodiments of the present invention, and it describes more concrete and detailed, but can not
Therefore the restriction to the scope of the claims of the present invention it is interpreted as.It should be pointed out that, for the person of ordinary skill of the art,
Without departing from the inventive concept of the premise, it is also possible to make some deformation and improvement, these broadly fall into the protection model of the present invention
Enclose.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.
Claims (10)
1. an anti-endometrial antibodies chemiluminescence immune detection reagent kit, it is characterised in that including: anti-endometrial antibodies
The coated carboxylated magnetic particle of recombiant protein and the chemiluminescent labels of human immunoglobulins's labelling.
Anti-endometrial antibodies chemiluminescence immune detection reagent kit the most according to claim 1, it is characterised in that described
In the coated carboxylated magnetic particle of anti-endometrial antibodies recombiant protein, described anti-endometrial antibodies recombiant protein is with described
The ratio of carboxylated magnetic particle is 1:25 ~ 35.
Anti-endometrial antibodies chemiluminescence immune detection reagent kit the most according to claim 1, it is characterised in that described
In the chemiluminescent labels of human immunoglobulins's labelling, described anti-endometrial antibodies recombiant protein and described chemiluminescence
The ratio of label is 50:1 ~ 10.
Anti-endometrial antibodies chemiluminescence immune detection reagent kit the most according to claim 1, it is characterised in that described
The particle diameter of carboxylated magnetic particle is 0.05 μm ~ 1 μm.
Anti-endometrial antibodies chemiluminescence immune detection reagent kit the most according to claim 1, it is characterised in that described
Chemiluminescent labels is luminol, different luminol, tris (bipyridine) ruthenium or acridinium ester.
Anti-endometrial antibodies chemiluminescence immune detection reagent kit the most according to claim 1, it is characterised in that also wrap
Including Chemoluminescent substrate, described Chemoluminescent substrate includes A liquid and B liquid.
Anti-endometrial antibodies 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.
Anti-endometrial antibodies chemiluminescence immune detection reagent kit the most according to claim 1, it is characterised in that also wrap
Include anti-endometrial antibodies calibration product.
Anti-endometrial antibodies chemiluminescence immune detection reagent kit the most according to claim 8, it is characterised in that described
Anti-endometrial antibodies calibration product are that concentration is respectively the anti-of 1U/L, 10U/L, 100U/L, 500U/L, 1000U/L and 2000U/L
The solution of AEA ELISA.
10. one kind according to the anti-endometrial antibodies chemiluminescence immune detection reagent kit according to any one of claim 1 ~ 9
Preparation 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 anti-endometrial antibodies recombiant protein, suspendible 2h ~ 10h under room temperature,
Magneto separate is resuspended with Tris buffer after removing supernatant, obtains the coated carboxylated magnetic of anti-endometrial antibodies recombiant protein micro-
Grain；And take human immunoglobulins, mix after adding carbonate buffer solution, mix after being subsequently adding chemiluminescent labels,
Under room temperature, remove impurity after lucifuge reaction 1h ~ 2h, obtains the chemiluminescent labels of human immunoglobulins's labelling.
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Cited By (1)
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| CN111856032A (en) * | 2020-07-21 | 2020-10-30 | 成都华西海圻医药科技有限公司 | Method for rapidly detecting concentration of anti-PD-L1 monoclonal antibody |
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| CN102043054A (en) * | 2010-10-26 | 2011-05-04 | 东北农业大学 | Cow recessive endometritis early diagnosis kit |
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| CN111856032A (en) * | 2020-07-21 | 2020-10-30 | 成都华西海圻医药科技有限公司 | Method for rapidly detecting concentration of anti-PD-L1 monoclonal antibody |
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