CN107561293A - Anti-hCG antibodies chemiluminescence immune detection reagent kit and preparation method thereof - Google Patents
Anti-hCG antibodies chemiluminescence immune detection reagent kit and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of anti-hCG antibodies chemiluminescence immune detection reagent kit and preparation method thereof, anti-hCG antibodies chemiluminescence immune detection reagent kit includes：The chemiluminescent labels of the magnetic particle of the coated carboxylated of anti-hCG antibodies recombinant protein and human immunoglobulins' mark.This anti-hCG antibodies chemiluminescence immune detection reagent kit can be using Full-automatic chemiluminescence immunoassay analysis meter as detection instrument, complete this anti-hCG antibodies chemiluminescence immune detection reagent kit of detection of anti-hCG antibodies, by experiment, its detection sensitivity reaches 1U/L, 10 times are at least improved relative to the detection method sensitivity of traditional anti-hCG antibodies, the accuracy of detection of this anti-hCG antibodies chemiluminescence immune detection reagent kit is higher.
Description
Technical field
The present invention relates to vitro detection field, more particularly to a kind of anti-hCG antibodies chemiluminescence to exempt from
Epidemic disease detection kit and preparation method thereof.
Background technology
Human chorionic gonadotrophin (HCG) is by a kind of glycoprotein of trophocyte's secretion of placenta, Neng Gouwei
The life-span of corpus luteum menstruatis is held, corpus luteum menstruatis increase is turned into corpus luteum graviditatis；It can also promote androgen by aromatization so as to convert
For estrogen, while progesterone is stimulated to be formed；Human Fetus Testis Testosterone Secretion is stimulated, promotes the differentiation of male's property；It is thin with parent thyroid gland
Born of the same parents' tsh receptor combines, and stimulates thyroid activity.Clinically, human chorionic gonadotrophin is used for women No-clay weak interbed or ovum
Steep sterility caused by dysmaturity, and a part for Assisted Reproductive Technology ART controlling excess stimulation protocol.It is also used for
The low promoting sexual gland hormone sexual disorder of male is treated, infertility caused by hypospermatogenesis and rush sexual gland pituitary function deficiency
Caused delayed puberty and cryptorchidism.
The common methods of clinical detection human chorionic gonadotrophin have enzyme linked immunosorbent assay, radiommunoassay at present
Method, enzyme-catalyzed chemical luminescence method, but all there is in place of some shortcomings for these methods.
First, enzyme linked immunosorbent assay
Enzyme linked immunosorbent assay (ELISA) is widely used, but there is also following weak points for this method：
(1) 12 × 8 types, 6 × 8 types, 8 × 12 types or the hole Special micro porous plate of complete plate 96 is used to be used as antigen coat apparatus and anti-
Container is answered, 12 batches, 6 batches, 8 batches or whole plate first use can only be divided into when in use, can not be carried out independent, single
The detection of person-portion；
(2) quantitative determination reagent type used is more, and each detection reagent will be contained with reagent bottle, and often be made
It is required for changing imbibition nozzle during with a kind of reagent to be filled into the micropore of microwell plate respectively, not only reagent bottle species is more, filling
The operation of reagent is also extremely cumbersome；
(3) lack the corresponding mark to detection information, just can only will appreciate that or know by checking the mark of kit external packing box
The product batch number and term of validity information of detection reagent are known, and the information known is uncontrolled in detection process, has very big
Randomness；
(4) detection reagent easily causes the cross pollution between various reagents and shadow in open space in detection process
Ring the accuracy of testing result；
(5) detection process uses hand-manipulated more, and the dosage of reagent or sample is not bery accurate, and operating process is extremely cumbersome and multiple
It is miscellaneous, bust easily occurs, the degree of accuracy of testing result and precision are poor；
(6) in the quantity configuration of detection project reagent set and using being above the person-portion of item number × 48/96, if necessary to examine
10 projects are surveyed, then the configuration of reagent and use number must be 10 × 48/96 person-portions, if only a sample needs detection 10
Individual different project, it is also desirable to configure the reagent of 10 × 48/96 person-portions, there is the shortcomings that inadequate economical rationality.
2nd, radio immunoassay
Radioimmunoassay method radioactive pollution is big, load procedure is cumbersome, complex operation, operating time are long, measurement result not
Stably, the deficiencies of reagent holding time is short, kit operating automation degree is low, necessary instrument is expensive part.
3rd, 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）With two kinds of alkaline phosphatase, but there is certain office
Sex-limited, horseradish peroxidase major defect is：Luminol, also can be by H in the case of the presence of no horseradish peroxidase2O2
It is luminous to aoxidize itself, background is of a relatively high, influences signal to noise ratio, and kinetics is complicated, and influence factor is more, is as a result not sufficiently stable,
The substrate for obtaining high sensitivity and plateau length is not easy.Alkaline phosphatase major defect is：Substrate reach plateau when
Between it is long, substrate cost is high, causes testing cost high, patient burden's weight.
Acridinium ester has detailed advantage as the direct chemiluminescence of label compared to enzyme-catalyzed chemical luminescence, main performance
：Reaction does not need 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, interference because
Element is few, and reagent stability is good, can be with two-point calibration, and system is simple, and exciting liquid cost is low, acridinium ester easily and protein bind, and
Photon yield is not reduced after connection.
The content of the invention
Based on this, it is necessary to provide a kind of detection sensitivity higher anti-hCG antibodies chemiluminescence
Immunity detection reagent and preparation method thereof.
A kind of anti-hCG antibodies chemiluminescence immune detection reagent kit, including：Anti-human chorionic promotees
The chemiluminescent labels of the magnetic particle of the coated carboxylated of gonadal hormone antibody recombinant protein and human immunoglobulins' mark.
In one embodiment, the magnetic of the coated carboxylated of anti-hCG antibodies recombinant protein is micro-
In grain, the ratio of the magnetic particle of the anti-hCG antibodies recombinant protein and the carboxylated is 1：25~35.
In one embodiment, it is described anti-human immune in the chemiluminescent labels of human immunoglobulins' mark
The ratio of globulin and the chemiluminescent labels is 50：1~10.
In one embodiment, the particle diameter of the magnetic particle of the carboxylated is 0.05 μm ~ 1 μm.
In one embodiment, the chemiluminescent labels are luminol, different luminol, tris (bipyridine) ruthenium or acridine
Ester.
In one embodiment, in addition to Chemoluminescent substrate, the Chemoluminescent substrate include A liquid and B liquid.
In one embodiment, the A liquid is H2O2Solution, the B liquid are NaOH solution.
In one embodiment, in addition to anti-hCG antibodies calibrate product.
In one embodiment, anti-hCG antibodies calibration product be concentration be respectively 1U/L,
The solution of 10U/L, 100U/L, 500U/L, 1000U/L and 2000U/L anti-hCG antibodies.
A kind of preparation method of above-mentioned anti-hCG antibodies chemiluminescence immune detection reagent kit, bag
Include following steps：
Take the suspension of the magnetic particle of carboxylated, Magneto separate to be resuspended after removing supernatant with MES buffer solutions, be subsequently added into the EDC aqueous solution,
The surface carboxyl groups of the magnetic particle of activated carboxyl, anti-hCG antibodies recombinant protein is subsequently added into, at room temperature
Suspension 2h ~ 10h, Magneto separate are resuspended after removing supernatant with Tris buffer solutions, obtain anti-hCG antibodies restructuring
The magnetic particle of the coated carboxylated of albumen；And
Human immunoglobulins are taken, are mixed after adding carbonate buffer solution, are mixed after then adding chemiluminescent labels, room temperature
Cleaned after lower lucifuge reaction 1h ~ 2h, obtain the chemiluminescent labels of human immunoglobulins' mark.
This anti-hCG antibodies chemiluminescence immune detection reagent kit can be sent out with full-automatic chemical
Light immunity analysis instrument is detection instrument, and this anti-human chorionic of detection for completing anti-hCG antibodies promotees sexual gland
Hormone antibody chemiluminescence immune detection reagent kit, by experiment, its detection sensitivity reaches 1U/L, relative to traditional anti-human
The detection method sensitivity of hcg antibody at least improves 10 times, and this anti-hCG resists
The accuracy of detection of body chemiluminescence immune detection reagent kit is higher.
Brief description of the drawings
Fig. 1 is the preparation of the anti-hCG antibodies chemiluminescence immune detection reagent kit of an embodiment
The flow chart of method；
Fig. 2 is the anti-hCG antibodies canonical plotting that embodiment 3 obtains.
Embodiment
In order to facilitate the understanding of the purposes, features and advantages of the present invention, it is below in conjunction with the accompanying drawings and specific real
Example is applied to be described in detail the embodiment of the present invention.Elaborate in the following description many details in order to
Fully understand the present invention.But the invention can be embodied in many other ways as described herein, art technology
Personnel can do similar improvement in the case of without prejudice to intension of the present invention, therefore the present invention is not by following public specific implementation
Limitation.
The anti-hCG antibodies chemiluminescence immune detection reagent kit of one embodiment, including：It is anti-human
The chemistry hair of the magnetic particle of the coated carboxylated of hcg antibody recombinant protein and human immunoglobulins' mark
Signal thing.
Preferably, in the magnetic particle of the coated carboxylated of anti-hCG antibodies recombinant protein, anti-human suede
The ratio of Chorionic Gonadotropin antibody recombinant protein and the magnetic particle of carboxylated is 1：25~35.
Preferably, in the chemiluminescent labels of human immunoglobulins' mark, human immunoglobulins and chemiluminescence
The ratio of label is 50：1~10.
Preferably, the particle diameter of the magnetic particle of carboxylated 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-hCG antibodies chemiluminescence immune detection reagent kit is also
Including Chemoluminescent substrate.
Chemoluminescent substrate includes A liquid and B liquid.A liquid can be H2O2Solution, B liquid can be NaOH solution.
In the present embodiment, A liquid is the H that concentration is 0.1mol/L2O2Solution, B liquid are that the NaOH that concentration is 0.25mol/L is molten
Liquid.
In other examples, above-mentioned anti-hCG antibodies chemiluminescence immune detection reagent kit is also
Product are calibrated including anti-hCG antibodies.
Anti-hCG antibodies calibration product be concentration be respectively 1U/L, 10U/L, 100U/L, 500U/L,
The solution of 1000U/L and 2000U/L anti-hCG antibodies.
Specifically, anti-hCG antibodies calibration product can use standard items buffer solution by anti-human chorionic
It is respectively the anti-human of 1U/L, 10U/L, 100U/L, 500U/L, 1000U/L and 2000U/L that promoting sexual gland hormone antibody, which is configured to concentration,
The solution of hcg antibody.
This anti-hCG antibodies chemiluminescence immune detection reagent kit is used for anti-human chorionic rush property
During glandular hormone antibody test, product are calibrated to anti-hCG antibodies using Full-automatic chemiluminescence immunoassay analysis meter
Detected, draw standard curve, be built in computer software；Then actual sample is tested, sample is calculated according to sample luminous value
Concentration；Performance finally is carried out to anti-hCG antibodies automatic chemiluminescence immunoassay system（Sensitivity,
Linearly, precision, interference）Evaluation.
This anti-hCG antibodies chemiluminescence immune detection reagent kit can be sent out with full-automatic chemical
Light immunity analysis instrument is detection instrument, and this anti-human chorionic of detection for completing anti-hCG antibodies promotees sexual gland
Hormone antibody chemiluminescence immune detection reagent kit, by experiment, its detection sensitivity reaches 1U/L, relative to traditional anti-human
The detection method sensitivity of hcg antibody at least improves 10 times, and this anti-hCG resists
The accuracy of detection of body chemiluminescence immune detection reagent kit is higher.
In addition, this anti-hCG antibodies chemiluminescence immune detection reagent kit is also with following excellent
Point：
1st, acridinium ester is selected as marker material, and is applied to chemiluminescence immunoassay system, and the luminescence system is directly change
Learn and light, compared with traditional enzyme-catalyzed chemical luminescence, the reaction does not need the participation of enzyme, more cost-effective；
2nd, it is wide from the chemiluminescence immunoassay system range of linearity of acridinium ester, 1U/L ~ 1000U/L can be reached, and it is traditional
The inspection range of linearity of the detection method of anti-hCG antibodies is 20U/L ~ 1000U/L；
3rd, acridinium ester chemiluminescent immunoassay system repeatability is high, and in batch and difference between batch is within 5%, and this is other chemistry
Luminescence immunoassay system is unapproachable；
4th, chemiluminescence immunoassay system has realized quantifying for sample, by built-in standard curve to test software, only needs to survey
Sample originally can directly obtain the concentration value of sample；
5th, chemiluminescence immunoassay system can realize full-automation, and the addition of reagent and sample has instrument completion entirely, operation
It is easier, reduce artificial error.
The preparation side of above-mentioned anti-hCG antibodies chemiluminescence immune detection reagent kit as shown in Figure 1
Method, comprise the following steps：
Take the suspension of the magnetic particle of carboxylated, Magneto separate to be resuspended after removing supernatant with MES buffer solutions, be subsequently added into the EDC aqueous solution,
The surface carboxyl groups of the magnetic particle of activated carboxyl, anti-hCG antibodies recombinant protein is subsequently added into, at room temperature
Suspension 2h ~ 10h, Magneto separate are resuspended after removing supernatant with Tris buffer solutions, obtain anti-hCG antibodies restructuring
The magnetic particle of the coated carboxylated of albumen.
MES（2- (N- morpholines) ethyl sulfonic acid）The concentration of buffer solution is 0.02M, pH 5.5.
The concentration of Tris buffer solutions is 0.1M and contains 2%BSA, pH 8.0.
EDC（1- ethyls -3- (3- dimethyl aminopropyls)-carbodiimides）The concentration of the aqueous solution is 10mg/mL ~ 20mg/
The ratio of mL, EDC and the magnetic particle of carboxylated is 0.05：0.1~1.
Preferably, in the magnetic particle of the coated carboxylated of anti-hCG antibodies recombinant protein, anti-human suede
The ratio of Chorionic Gonadotropin antibody recombinant protein and the magnetic particle of carboxylated is 1：25~35.
Preferably, the particle diameter of the magnetic particle of carboxylated is 0.05 μm ~ 1 μm.
Human immunoglobulins are taken, are mixed after adding carbonate buffer solution, are mixed after then adding chemiluminescent labels,
Cleaned at room temperature after lucifuge reaction 1h ~ 2h, obtain the chemiluminescent labels of human immunoglobulins' mark.
Carbonate buffer solution concentration is 0.1M, and pH is 9.0 ~ 9.5,
The operation of removal of impurities is centrifugation desalting column desalination, and concrete operations are：Pure water and TBS buffer solutions are first used respectively（40 mM
Tris-HCl, 0.5% BSA, 1% NaCl, pH 8.0）Processing centrifugation desalting column, it is eventually adding obtained anti-human chorionic rush property
The solution of the magnetic particle of the coated carboxylated of glandular hormone antibody recombinant protein, finally collect the liquid in centrifuge tube.
Preferably, in the chemiluminescent labels of human immunoglobulins' mark, anti-hCG antibodies
The ratio of recombinant protein and 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 magnetic particle of the obtained coated carboxylated of anti-hCG antibodies recombinant protein and anti-human immune
The chemiluminescent labels combination of globulin mark can obtain above-mentioned anti-hCG antibodies chemiluminescence and exempt from
Epidemic disease detection kit.
This anti-hCG antibodies chemiluminescence immune detection reagent kit is when in use, it is also necessary to chemistry
Luminous substrate liquid and anti-hCG antibodies calibration product.
Chemoluminescent substrate and anti-hCG antibodies calibration product can voluntarily be prepared to 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 is the H that concentration is 0.1mol/L2O2Solution, B liquid are that the NaOH that concentration is 0.25mol/L is molten
Liquid.
Specifically, anti-hCG antibodies calibration product can use standard items buffer solution by anti-human chorionic
It is respectively the anti-human of 1U/L, 10U/L, 100U/L, 500U/L, 1000U/L and 2000U/L that promoting sexual gland hormone antibody, which is configured to concentration,
The solution of hcg antibody.
The preparation method of this anti-hCG antibodies chemiluminescence immune detection reagent kit is simple and convenient,
The detection sensitivity of obtained anti-hCG antibodies chemiluminescence immune detection reagent kit is higher, has good
Application prospect.
It is specific embodiment below.
Embodiment 1：The preparation of anti-hCG antibodies chemiluminescence immune detection reagent kit
（1）The preparation of the magnetic particle of the coated carboxylated of anti-hCG antibodies recombinant protein：
Take the magnetic particle containing the carboxylated that 50mg particle diameters are 0.05 μm ~ 1 μm（MagnaBind21353）Suspension, Magneto separate are gone
Supernatant, it is resuspended with 0.02 M, pH for 5.5 MES buffer solutions, adds the EDC aqueous solution for the 10mg/mL that 1mL is newly configured, activate magnetic
Bead surface carboxyl, add 4mg anti-hCG antibodies recombinant proteins（Biorbyt, article No. orb48780）, room temperature
Lower suspension 6h, Magneto separate, supernatant is removed, with the 0.1M containing 2%BSA, the Tris buffer solutions that pH is 8.0 are resuspended to 1mg/mL, obtained
The magnetic particle of the coated carboxylated of anti-hCG antibodies recombinant protein, every bottle of 5mL packing be stored in 4 DEG C it is standby
With.
（2）The preparation of the acridinium ester of human immunoglobulins' mark：
The human immunoglobulins that 50 μ L concentration are 25mg/mL are taken, it is the carbonic acid that 0.1M, pH are 9.0 ~ 9.5 to add 150 μ L concentration
Salt buffer, mix, then add the acridine ester solution that 1.5 μ L concentration are 5mg/mL and mix, lucifuge is reacted at room temperature, after 1.5h
Take out, centrifuge desalting column desalting processing with 2mL zeba, carried out respectively with pure water and TBS buffer solutions first in desalination processes
Processing, the acridine ester solution for the human immunoglobulins' mark for being eventually adding to obtain, liquid in centrifuge tube is collected to preserving pipe
Obtain the acridinium ester of human immunoglobulins' mark, every bottle of 5mL packing be stored in 4 DEG C it is standby.
（3）Anti-hCG antibodies calibrate the preparation of product：
With standard items buffer solution（40 mM Tris-HCl, 0.5% BSA, 1% NaCl, pH 8.0）Anti-human chorionic is promoted into sexual gland
It is 0U/L, 10U/L, 100U/L, 500U/L, 1000U/L and 2000U/L that hormone antibody, which is configured to concentration, and every bottle of 0.5 mL is dispensed
Lyophilized, 4 DEG C save backup.
Embodiment 2：Anti-hCG antibodies chemical luminous immune detection method
With Full-automatic chemiluminescence immunoassay analysis meter（YHLO, article No. iFlash3000）To detect instrument, between methodology pattern is
Immunization is connect, i.e. instrument sequentially adds 50 μ L sample, 50 μ L anti-hCG antibodies recombinant protein bag
The anti-hCG antibodies treatment fluid of the magnetic particle of the carboxylated of quilt and 50 μ L, after reacting 20 min, then add
50 μ L human immunoglobulins' acridinium ester, after reacting 20 min, Magneto separate being carried out, reactant mixture is sent into darkroom by instrument,
Sequentially add luminous substrate A liquid（H2O2）And B liquid（NaOH）Luminescence-producing reaction is carried out, finally records luminous value.
Embodiment 3：Anti-hCG antibodies chemiluminescence immune detection reagent kit performance evaluation
Anti-hCG antibodies calibration product are detected using the method in embodiment 2, obtain drafting standard
Curve is as shown in Figure 2.
Then to then testing actual sample, concentration of specimens is calculated according to sample luminous value.
The detection of sensitivity：
With reference to CLSI EP17-A file recommendation experimental programs, anti-hCG antibodies chemiluminescence immunoassay is calculated
The sensitivity of detection kit, the sensitivity tried to achieve are 1U/L.
Linear detection：
It is that 1U/L, 10U/L, 100U/L, 500U/L, 1000U/L and 2000U/L standard items do linear analysis to concentration, calculates line
Property coefficient correlation, r=0.9996, in addition, the range of linearity of the kit to anti-hCG antibodies sample detection
For 1U/L ~ 1000U/L.
Precision determines：
It is two anti-hCG antibodies samples of 50U/L and 500U/L to take concentration, and each each concentration of sample is each
Do 3 it is parallel, detected, calculated in kit batch and difference between batch with three batches of kits, the results showed that in the kit batch and
Difference between batch is respectively less than 5%.
Interference is tested：
Taking pooled serum to add chaff interference respectively includes：With reference to bilirubin, unconjugated bilirubin, hemoglobin, ascorbic acid, glycerine
Ester, adding proportion is according to 1：20 are carried out, and are determined pooled serum respectively and be with the addition of the measured value of pooled serum after various chaff interferences,
Deviation therebetween is calculated, with ± 10% for tolerance interval.As a result show, interference reaches NCCLS files-designated
Standard, the accurate evaluation available for clinical labororatory's anti-hCG antibodies situation.
The contrast experiment of embodiment 4, anti-hCG antibodies chemiluminescence immune detection reagent kit
The anti-human chorionic that concentration is 0,50U/L is promoted with chemical luminescence detection method and traditional enzyme linked immunosorbent assay respectively
Gonadal hormone antibody samples are detected, and two methods detection sensitivity is compared, and data are as shown in the table：
As can be seen from the above table, the sensitivity of chemical luminescence detection method improves more than 10 times compared with enzyme linked immunosorbent assay.
Embodiment described above only expresses the several embodiments of the present invention, and its description is more specific and detailed, but can not
Therefore it is interpreted as the limitation to the scope of the claims of the present invention.It should be pointed out that for the person of ordinary skill of the art,
Without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to the protection model of the present invention
Enclose.Therefore, the protection domain of patent of the present invention should be determined by the appended claims.
Claims (10)
- A kind of 1. anti-hCG antibodies chemiluminescence immune detection reagent kit, it is characterised in that including：It is anti-human The chemistry hair of the magnetic particle of the coated carboxylated of hcg antibody recombinant protein and human immunoglobulins' mark Signal thing.
- 2. anti-hCG antibodies chemiluminescence immune detection reagent kit according to claim 1, it is special Sign is, in the magnetic particle of the coated carboxylated of anti-hCG antibodies recombinant protein, the anti-human suede The ratio of Chorionic Gonadotropin antibody recombinant protein and the magnetic particle of the carboxylated is 1：25~35.
- 3. anti-hCG antibodies chemiluminescence immune detection reagent kit according to claim 1, it is special Sign is, in the chemiluminescent labels of human immunoglobulins' mark, the anti-hCG antibodies The ratio of recombinant protein and the chemiluminescent labels is 50：1~10.
- 4. anti-hCG antibodies chemiluminescence immune detection reagent kit according to claim 1, it is special Sign is that the particle diameter of the magnetic particle of the carboxylated is 0.05 μm ~ 1 μm.
- 5. anti-hCG antibodies chemiluminescence immune detection reagent kit according to claim 1, it is special Sign is that the chemiluminescent labels are luminol, different luminol, tris (bipyridine) ruthenium or acridinium ester.
- 6. anti-hCG antibodies chemiluminescence immune detection reagent kit according to claim 1, it is special Sign is, in addition to Chemoluminescent substrate, and the Chemoluminescent substrate includes A liquid and B liquid.
- 7. anti-hCG antibodies chemiluminescence immune detection reagent kit according to claim 6, it is special Sign is that the A liquid is H2O2Solution, the B liquid are NaOH solution.
- 8. anti-hCG antibodies chemiluminescence immune detection reagent kit according to claim 1, it is special Sign is, in addition to anti-hCG antibodies calibration product.
- 9. anti-hCG antibodies chemiluminescence immune detection reagent kit according to claim 8, it is special Sign is that the anti-hCG antibodies calibration product are that concentration is respectively 1U/L, 10U/L, 100U/L, 500U/ L, the solution of 1000U/L and 2000U/L anti-hCG antibodies.
- A kind of 10. anti-hCG antibodies chemiluminescence immunoassay according to any one of claim 1 ~ 9 The preparation method of detection kit, it is characterised in that comprise the following steps：Take the suspension of the magnetic particle of carboxylated, Magneto separate to be resuspended after removing supernatant with MES buffer solutions, be subsequently added into the EDC aqueous solution, The surface carboxyl groups of the magnetic particle of activated carboxyl, anti-hCG antibodies recombinant protein is subsequently added into, at room temperature Suspension 2h ~ 10h, Magneto separate are resuspended after removing supernatant with Tris buffer solutions, obtain anti-hCG antibodies restructuring The magnetic particle of the coated carboxylated of albumen；And human immunoglobulins are taken, mix after adding carbonate buffer solution, then add Mix after chemiluminescent labels, cleaned at room temperature after lucifuge reaction 1h ~ 2h, obtain the chemistry hair of human immunoglobulins' mark Signal thing.
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Application publication date: 20180109 |