CN107782889A - A kind of Ciclosporin A immunologic function test reagent and its preparation and detection method - Google Patents
A kind of Ciclosporin A immunologic function test reagent and its preparation and detection method Download PDFInfo
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- CN107782889A CN107782889A CN201711388780.1A CN201711388780A CN107782889A CN 107782889 A CN107782889 A CN 107782889A CN 201711388780 A CN201711388780 A CN 201711388780A CN 107782889 A CN107782889 A CN 107782889A
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- ciclosporin
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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
Abstract
The invention discloses a kind of Ciclosporin A immunologic function test reagent and its preparation and detection method.Including：Enzyme mark Ciclosporin A, the indicator for detecting Ciclosporin A enzyme-labeled antibody Ciclosporin A compound；Above-mentioned enzyme mark Ciclosporin A is formed by Ciclosporin A and glucose dehydrogenase coupling.The Ciclosporin A immunologic function test reagent of the present invention can accurately and quickly determine Ciclosporin A content in the samples such as blood of human body.Compared with the existing detection reagent of in the market, detection reagent of the present invention has the advantages that fast and easy, high sensitivity, high specificity, quantitative accurate, is advantageous to promoting the use of for clinic.
Description
Technical field
The present invention relates to field of biological detection, specifically a kind of Ciclosporin A immunologic function test reagent and its preparation and detection
Method.
Background technology
Ciclosporin A (Cyclosporin, CsA) is a kind of small molecule ring type polypeptide chemical combination for including 11 amino acid
Thing.CsA is the rejection potent immunosuppressant inhibitor of current clinical conventional allogeneic organ or tissue transplantation, to cell and
Humoral immunity has compared with high selectivity inhibitory action, is a kind of lymphocyte activator inhibitor.The main suppression complementary lymphs of T are thin
Born of the same parents, also have to T wound property cell and acted on compared with high inhibition.CsA is using increasingly extensive, but it has complicated pharmacokinetics and pharmacodynamics special
Point, toxic side effect is more and incidence is higher, and individual difference is big, and toxicity is not easy to distinguish with rejection, and monitoring difficulty is big, its
Blood concentration is too high to cause toxicity, especially to liver, the toxicity of kidney, therefore carry out blood concentration prison over the course for the treatment of
Survey and pharmacokinetic, the survival rate to improving transplant organ, the generation for reducing toxic reaction and rejection have weight
The clinical meaning wanted.
The common method of detection Ciclosporin A has at present：Efficient liquid phase chromatographic analysis, radiommunoassay（RIA）With it is glimmering
Light polarization method（FPIA）Deng.Wherein, efficient liquid phase chromatographic analysis is time-consuming and is not easy to automate；It is longer the time required to RIA methods, detection
Unstable result, and radioactive pollution danger be present.FPIA methods need expensive instrument and equipment and veteran operating personnel.
For homogeneous enzyme immunoassay detection method, its advantage is the method that the present invention uses：Easy to operate, quick, high sensitivity, standard
Really property is good, is suitable for automating, and is widely used, and with automatic clinical chemistry analyzer to small-molecule substance and macromolecular substances all
Energy high flux quickly determines.
The content of the invention
It is accurate it is an object of the invention to solve Ciclosporin A detection process complex operation in the prior art and measure
The problem of low is spent, the invention provides a kind of quick, high sensitivity, accurately detects Ciclosporin A content in sample to be tested
Ciclosporin A homogeneous enzyme immunoassay detection reagent and preparation method thereof.
To achieve the above object, the present invention provides following technical scheme：
A kind of Ciclosporin A immunologic function test reagent and its preparation and detection method, it is characterised in that：Enzyme mark Ciclosporin A, it is used for
Detect the indicator of Ciclosporin A antibody-enzyme mark Ciclosporin A compound；Above-mentioned enzyme mark Ciclosporin A is by Ciclosporin A
Formed with glucose dehydrogenase coupling.
As the further scheme of the present invention, the indicator is selected from enzymatic reagent, including：The bottom of enzyme mark conjugate and enzyme
Thing；Above-mentioned enzyme mark conjugate includes glucose dehydrogenase-Ciclosporin A conjugate；The substrate of above-mentioned enzyme is glucose.
As the further scheme of the present invention, a kind of described Ciclosporin A immunologic function test reagent and preparation method thereof, its
It is characterised by, comprises the following steps：
（1）The preparation of glucose dehydrogenase-Ciclosporin A conjugate：Glucose dehydrogenase（GDH）It is coupled with Ciclosporin A, it is pure
Change the enzyme-labelled antigen of coupling；
（2）The preparation of Ciclosporin A homogeneous enzyme immunoassay detection reagent：
The preparation of reagent 1：Mixed by Ciclosporin A antibody and homogeneous zymolyte；
The preparation of reagent 2：Mixed by glucose dehydrogenase-antigen conjugates with phosphate buffer.
As the further scheme of the present invention, a kind of preparation method of described Ciclosporin A immunologic function test reagent, it is special
Sign is, described step（1）Detailed process is：
1）Glucose dehydrogenase（GDH）With Ciclosporin A（CsA）Coupling
A. 100-400 mg Ciclosporin As, 20-100 mg 4- benzoylbenzoic acids are accurately weighed（BBa）, load quartz cuvette
In ware, the 5-20 mL tert-butyl alcohols, ultrasonic dissolution are added；
B. ultraviolet light 2-5 hours, reaction is abundant, is stored at room temperature into freeze-dried powder；
C. 2-10 mg GDH are accurately weighed to be dissolved in 0.1-1 mL PBS solution, take above-mentioned powder（CsA-BBa）10-50 mg
It is dissolved in 0.5-2 mL dimethylformamide（DMF）In solution, under the stirring of 25 DEG C of constant temperature by CsA-BBa be added dropwise to dissolved with
In GDH PBS solution；
D. the pH for adjusting solution is maintained at 6.0-8.0, then 0.5% carbodiimides is added dropwise（EDC）50-200 μ L, 4 DEG C
It is stirred overnight；
2）Purify the enzyme-labelled antigen of coupling
By the enzyme-labelled antigen of G-25 gel chromatography columns purifying coupling, glucose dehydrogenase-Ciclosporin A conjugate is obtained, in
Stored at 2-8 DEG C.
As the further scheme of the present invention, a kind of preparation method of described Ciclosporin A immunologic function test reagent, it is special
Sign is, step（2）Detailed process it is as follows：
The preparation of reagent 1：By the NADH NAD of 2-5 g oxidation state, 0.5-3 g glucose 0.5-2L
Homogeneous zymolyte is made in phosphate buffer dissolving；Ciclosporin A antibody is added in above-mentioned homogeneous zymolyte, antibody with it is homogeneous
The volume ratio of zymolyte is 1:100~1:10000；
The preparation of reagent 2：The glucose dehydrogenase of preparation-Ciclosporin A conjugate is added in phosphate buffer, above-mentioned idol
The volume ratio for joining thing and phosphate buffer is 1:100~1:10000.
Exist as the further scheme of the present invention, the detection method of described Ciclosporin A immunologic function test reagent, its feature
In comprising the following steps：
1）Sample to be tested is contacted with Ciclosporin A antibody；
2）According to the combination situation of enzyme mark Ciclosporin A in sample to be tested and Ciclosporin A antibody, sample is judged using indicator
The content of Ciclosporin A in this；The sample to be tested is serum, blood plasma, saliva or urine.
The principle of the present invention is that antigen is combined into enzyme-labelled antigen with enzyme, retains the bioactivity of antigen and enzyme, when enzyme mark resists
After former and antibody binding, zymoprotein and antibody close contact on antigen molecule, the activated centre of enzyme is set to be affected, the work of enzyme
Property is suppressed.Antigen, enzyme-labelled antigen during measure in sample are combined with antibody competition, and the antigenic content in sample is higher,
Its OD value is higher after adding substrate.
The advantage of the invention is that：The Ciclosporin A immunologic function test reagent of the present invention can accurately and quickly determine human body
Ciclosporin A content in the samples such as blood.Compared with the existing detection reagent of in the market, detection reagent of the present invention has convenient fast
Speed, high sensitivity, high specificity, it is quantitative accurate the advantages that, be advantageous to promoting the use of for clinic.
Brief description of the drawings
Fig. 1 is Ciclosporin A homogeneous enzyme immunoassay reaction calibration graph.
Fig. 2 is Ciclosporin A homogeneous enzyme immunoassay range of linearity figure.
Fig. 3 is the correlation analysis figure of the chemoluminescence method testing result of Ciclosporin A homogeneous enzyme immunoassay and NPD projects.
Embodiment
The invention provides a kind of Ciclosporin A immunologic function test reagent and its preparation and detection method, to make mesh of the present invention
, technical scheme and effect it is clearer, clear and definite, the present invention is described in detail below.
The invention provides a kind of Ciclosporin A immunologic function test reagent and its preparation and detection method.Including：Enzyme mark ring spore
Mycin A, the indicator for detecting Ciclosporin A antibody-enzyme mark Ciclosporin A compound；Above-mentioned enzyme mark Ciclosporin A by
Ciclosporin A and glucose dehydrogenase coupling form.
Signified " Ciclosporin A " refers not only to complete Ciclosporin A molecule in the present invention, also includes retaining complete resist
The Ciclosporin A segment or derivative of former specific binding capacity.
A kind of Ciclosporin A homogeneous enzyme immunoassay detection reagent, including：Enzyme mark Ciclosporin A, for detecting Ciclosporin A
The indicator of antibody-enzyme mark Ciclosporin A compound.Indicator is selected from enzymatic reagent, radio isotope reagent, fluorescence examination
Agent and chemical illuminating reagent.Preferably, indicator is enzymatic reagent, including：The substrate of enzyme mark conjugate and enzyme.Wherein, enzyme mark
Conjugate includes glucose dehydrogenase-antigen conjugates, and it can be obtained by chemical synthesis process.
The application method of above-mentioned Ciclosporin A immunologic function test reagent, comprises the following steps：
1）Sample to be tested is contacted with Ciclosporin A antibody；
2）According to the combination situation of enzyme mark Ciclosporin A in sample to be tested and Ciclosporin A antibody, sample is judged using indicator
The content of Ciclosporin A in this；The sample to be tested is serum, blood plasma, saliva or urine etc..Preferably, sample to be tested is blood
Clear or blood plasma.
Below by specific embodiment, the present invention is described in detail.
Embodiment one：The preparation of glucose dehydrogenase-antigen conjugates
1）Glucose dehydrogenase（GDH）With Ciclosporin A（CsA）Coupling
A. 240 mg Ciclosporin As, 50 mg 4- benzoylbenzoic acids are accurately weighed（BBa）, it is fitted into quartz colorimetric utensil, adds
Enter the 10 mL tert-butyl alcohols, ultrasonic dissolution；
B. ultraviolet light 3 hours, reaction is abundant, is stored at room temperature into freeze-dried powder；
C. 4.5 mg GDH are accurately weighed to be dissolved in 0.5 mL 10mM pH 7.4 PBS solution, take above-mentioned powder（CsA-
BBa）15 mg are dissolved in 0.7 mL dimethylformamide（DMF）In solution, CsA-BBa is added dropwise under 25 DEG C of stirrings of constant temperature
Into the PBS solution dissolved with GDH；
D. regulation pH value of solution is maintained at 6.0-8.0, then 0.5% carbodiimides is added dropwise（EDC）120 μ L, 4 DEG C of stirrings
Overnight.
2）Purify the enzyme-labelled antigen of coupling
By the enzyme-labelled antigen of G-25 gel chromatography columns purifying coupling, glucose dehydrogenase-Ciclosporin A conjugate is obtained, in
Stored at 2-8 DEG C.
Embodiment two：The preparation of Ciclosporin A homogeneous enzyme immunoassay detection reagent
Ciclosporin A homogeneous enzyme immunoassay detection reagent, including：Enzyme mark Ciclosporin A, for detecting Ciclosporin A antibody-enzyme mark
The indicator of Ciclosporin A compound.Indicator is selected from enzymatic reagent, radio isotope reagent, fluorometric reagent and chemistry
Luminescence reagent.Preferably, indicator is enzymatic reagent, including：The substrate of enzyme mark conjugate and enzyme.Wherein, enzyme mark conjugate bag
Glucose dehydrogenase-antigen conjugates are included, it can be obtained by chemical synthesis process.
Ciclosporin A homogeneous enzyme immunoassay detection reagent before the use, in order to avoid the enzyme mark conjugate in indicator
Reacted with the substrate of enzyme, the substrate of enzyme mark conjugate and enzyme is separated, therefore Ciclosporin A homogeneous enzyme immunoassay is examined
Test agent includes two kinds of reagents being provided separately, specific as follows：
1. the preparation of reagent 1：By 3.588g（10mM）NADH NAD, 1.802g of oxidation state（10mM）
Homogeneous zymolyte is made with mM, the pH 8.0 of 1L 50 phosphate buffer dissolving in glucose；Ciclosporin A antibody is added to
State in homogeneous zymolyte, the volume ratio of antibody and homogeneous zymolyte is 1:100~1:10000, it is specific in the present embodiment to compare
Example is 1:600.
2. the preparation of reagent 2：The glucose dehydrogenase of preparation-Ciclosporin A conjugate is added to 50mM, pH8.0 phosphorus
In phthalate buffer, the volume ratio of above-mentioned conjugate and phosphate buffer is 1:100~1:10000, have in the present embodiment
The ratio of body is 1:2000.
The application method of above-mentioned Ciclosporin A homogeneous enzyme immunoassay detection reagent, comprises the following steps：
1）Sample to be tested is contacted with Ciclosporin A antibody；
2）According to the combination situation of enzyme mark Ciclosporin A in sample to be tested and Ciclosporin A antibody, sample is judged using indicator
The content of Ciclosporin A in this；
Specifically, sample to be tested is added in reagent 1 during detection, the Ciclosporin A in sample to be tested and the ring spore in reagent 1 are mould
Plain A antibody is specifically bound, and generates anti-Ciclosporin A antibody-Ciclosporin A compound；Reagent 2 is added, is now tried
Glucose dehydrogenase-Ciclosporin A conjugate in agent 2 is mixed with the substrate of the enzyme in reagent 1, contacted, and enzymatic reaction occurs,
The indicator of detection Ciclosporin A antibody-enzyme mark Ciclosporin A compound is formed, indicator is according to sample to be tested middle ring
The combination situation of p0-357 A and above-mentioned Ciclosporin A antibody judges the content of Ciclosporin A in sample to be tested.
Due to the Ciclosporin A competitive binding Ciclosporin A in glucose dehydrogenase-antigen conjugates and sample to be tested
Antibody, so, the amount of Ciclosporin A is more in sample to be tested, the glucose dehydrogenase dissociated in homogeneous enzyme solutions-antigen coupling
The amount of thing is more, and enzymatic reaction is faster, causes OD340 Rise.
Above-mentioned sample to be tested is physiology sample, such as serum, blood plasma, urine, saliva etc., as a kind of preferable scheme,
Above-mentioned sample to be tested is serum or blood plasma.
Embodiment three：Ciclosporin A homogeneous enzyme immunoassay detection reagent reacts calibration curve.
1）CsA calibration objects are prepared：Commercially available people's Ciclosporin A recombinant protein is dissolved in the solution of similar human serum matrix
（NaCl 0.9%, BSA 0.2%, NaN3 0.1%, Tris-HCl pH 7.4）In, the calibration objects of various concentrations is made.With Shenzhen
NPD projects biomedical engineering Co., Ltd Ciclosporin A calibration object is primary standard, using its Ciclosporin A kit to not
Calibration object with concentration detects 10 times respectively, obtains average, obtains the concentration of Ciclosporin A calibration object：25,50,100,200,
400,800 ng/mL.
2）Biochemical Analyzer detects：So that Hitachi 7170 operates as an example：Measure wavelength is 340nm, takes various concentrations respectively
Calibration object solution（20 μL）, add Ciclosporin A R1Reagent（160 μL）, mix, add Ciclosporin A R2Reagent（40
μL）, after mixing, determine the OD of different time points340Light absorption value, calculate reaction rate during different calibration object concentration, actual behaviour
The constantly volume ratio of adjustment reagent 1 and reagent 2 is needed during work, while adjusts light-metering point, finally draws comparatively ideal reaction
Canonical plotting, often pipe replication 3 times, using 3 absorbance difference Δ A measured of each calibration pipe average value as ordinate,
Corresponding calibration object concentration is abscissa, draws " concentration-absorbance difference " calibration curve（See Fig. 1）.
Test serum or plasma sample are taken, is measured in the same method the absorbance difference of sample, substitutes into calibration curve, you can calculate
The content of Ciclosporin A in sample to be tested.If the concentration of Ciclosporin A exceeds calibration curve scope in serum or blood plasma, need
Detected again after being diluted to sample to ensure the accuracy of testing result.
This detection reagent is applicable not only to Hitachi 7170, applies also for semi-automatic, the full-automatic life of other brands and model
Change analyzer, design parameter can be adjusted according to instrument.
Example IV：The range of linearity determines
With the Ciclosporin A high concentration sample close to the range of linearity upper limit（780.8 ng/mL）, with above-mentioned similar human serum matrix
Solution by its by 1/2,1/4,1/8,1/16,1/32,1/64 dilution, be configured to 6 diluted concentrations altogether（xi）Solution, use institute
State Biochemical Analyzer detection method and determine each diluted sample concentration.Each diluted concentration is tested 3 times, and it is dense to obtain each dilution respectively
Spend the average of testing result（yi）.With diluted concentration（xi）For independent variable, to determine average（yi）Linear return is obtained for dependent variable
Return equation, according to formula（1）The correlation coefficient r of linear regression is calculated, it is y=0.9985x-0.1818 as a result to show regression equation,
Correlation coefficient r=0.9999, show reagent of the present invention good relationship in the 22.5 ng/mL -780.8 ng/mL ranges of linearity
（See Fig. 2）.
Embodiment five：Correlation analysis
To 85 clinical samples respectively using the chemoluminescence method of NPD projects and the homogeneous enzyme immunoassay method of the present invention respectively to sample
Replication 2 times, calculates average value respectively, and linear regression analysis is carried out to 85 pattern detection results, calculates two kinds of reagent inspections
Survey the coefficient correlation of result.
As a result show, correlation coefficient r=0.9961 of two kinds of kit testing results, equation of linear regression is
y=0.9879x+0.5373（See Fig. 3）, show Ciclosporin A homogeneous enzyme immunoassay method detection reagent of the present invention and commercially available new production
The chemoluminescence method detection reagent of industry has good uniformity.
Because the detection process of the present invention is completed by instrument is full-automatic, so to the less demanding of testing staff, easily
In realizing and promote the use of.
It should be noted that obviously the invention is not restricted to the details of above-mentioned one exemplary embodiment, the scope of the present invention is by institute
Attached claim rather than described above limit, it is intended that will fall in the implication and scope of the equivalency of claim
All changes are included in the scope of patent protection of the present invention.
In addition, above-described is only the preferred embodiments of the invention, for the technical staff in this technology neck city,
Under the premise without departing from the principles of the invention, some modifications and adaptations can also be done, these improved adjustment also should be regarded as this
The protection domain of invention.
Claims (7)
1. a kind of Ciclosporin A immunologic function test reagent and its preparation and detection method, it is characterised in that：Enzyme mark Ciclosporin A, use
In the indicator of detection Ciclosporin A antibody-enzyme mark Ciclosporin A compound.
2. the immunologic function test reagent of Ciclosporin A according to claim 1, it is characterised in that：The enzyme mark Ciclosporin A
Formed by Ciclosporin A and glucose dehydrogenase coupling.
3. Ciclosporin A immunologic function test reagent according to claim 1, it is characterised in that：The indicator is selected from enzyme
Reagent, including：The substrate of enzyme mark conjugate and enzyme；Above-mentioned enzyme mark conjugate includes glucose dehydrogenase-Ciclosporin A coupling
Thing；The substrate of above-mentioned enzyme is glucose.
4. a kind of Ciclosporin A immunologic function test reagent and preparation method thereof, it is characterised in that comprise the following steps：
（1）The preparation of glucose dehydrogenase-Ciclosporin A conjugate：Glucose dehydrogenase（GDH）It is coupled with Ciclosporin A, it is pure
Change the enzyme-labelled antigen of coupling；
（2）The preparation of Ciclosporin A homogeneous enzyme immunoassay detection reagent：
The preparation of reagent 1：Mixed by Ciclosporin A antibody and homogeneous zymolyte；
The preparation of reagent 2：Mixed by glucose dehydrogenase-Ciclosporin A conjugate with phosphate buffer.
5. the preparation method of a kind of Ciclosporin A immunologic function test reagent according to claim 5, it is characterised in that described
The step of（1）Detailed process is：
1）Glucose dehydrogenase（GDH）With Ciclosporin A（CsA）Coupling
A. 100-400 mg Ciclosporin As, 20-100 mg 4- benzoylbenzoic acids are accurately weighed（BBa）, load quartz cuvette
In ware, the 5-20 mL tert-butyl alcohols, ultrasonic dissolution are added；
B. ultraviolet light 2-5 hours, reaction is abundant, is stored at room temperature into freeze-dried powder；
C. 2-10 mg GDH are accurately weighed to be dissolved in 0.1-1 mL PBS solution, take above-mentioned powder（CsA-BBa）10-50 mg
It is dissolved in 0.5-2 mL dimethylformamide（DMF）In solution, under the stirring of 25 DEG C of constant temperature by CsA-BBa be added dropwise to dissolved with
In GDH PBS solution；
D. the pH for adjusting solution is maintained at 6.0-8.0, then 0.5% carbodiimides is added dropwise（EDC）50-200 μ L, 4 DEG C
It is stirred overnight；
2）Purify the enzyme-labelled antigen of coupling
By the enzyme-labelled antigen of G-25 gel chromatography columns purifying coupling, glucose dehydrogenase-Ciclosporin A conjugate is obtained, in
Stored at 2-8 DEG C.
A kind of 6. preparation method of Ciclosporin A immunologic function test reagent according to claim 5, it is characterised in that step
（2）Detailed process it is as follows：
The preparation of reagent 1：By the NADH NAD of 2-5 g oxidation state, 0.5-3 g glucose 0.5-2 L
Homogeneous zymolyte is made in phosphate buffer dissolving；Ciclosporin A antibody is added in above-mentioned homogeneous zymolyte, antibody with it is homogeneous
The volume ratio of zymolyte is 1:100~1:10000；
The preparation of reagent 2：The glucose dehydrogenase of preparation-Ciclosporin A conjugate is added in phosphate buffer, above-mentioned idol
The volume ratio for joining thing and phosphate buffer is 1:100~1:10000.
7. using the detection method of the Ciclosporin A immunologic function test reagent described in any one of Claims 1-4 4, its feature exists
In comprising the following steps：
1）Sample to be tested is contacted with Ciclosporin A antibody；
2）According to the combination situation of Ciclosporin A in sample to be tested and Ciclosporin A antibody, using in indicator judgement sample
The content of Ciclosporin A；The sample to be tested is serum, blood plasma, saliva or urine.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101048660A (en) * | 2004-08-27 | 2007-10-03 | 灵芝国际股份有限公司 | Homogeneous enzyme immunoassay for oral fluid |
| CN103421187A (en) * | 2012-05-18 | 2013-12-04 | 华中科技大学同济医学院附属协和医院 | Method for preparing cyclosporin A holoantigen |
| CN104764883A (en) * | 2015-03-31 | 2015-07-08 | 上海云泽生物科技有限公司 | Immunoassay method and kit for detecting concentration of cyclosporine A |
| CN104788560A (en) * | 2015-05-16 | 2015-07-22 | 苏州博源医疗科技有限公司 | Cyclosporin A immunogen, anti-cyclosporin A specific antibody and cyclosporin A detection reagent |
-
2017
- 2017-12-22 CN CN201711388780.1A patent/CN107782889A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101048660A (en) * | 2004-08-27 | 2007-10-03 | 灵芝国际股份有限公司 | Homogeneous enzyme immunoassay for oral fluid |
| CN103421187A (en) * | 2012-05-18 | 2013-12-04 | 华中科技大学同济医学院附属协和医院 | Method for preparing cyclosporin A holoantigen |
| CN104764883A (en) * | 2015-03-31 | 2015-07-08 | 上海云泽生物科技有限公司 | Immunoassay method and kit for detecting concentration of cyclosporine A |
| CN104788560A (en) * | 2015-05-16 | 2015-07-22 | 苏州博源医疗科技有限公司 | Cyclosporin A immunogen, anti-cyclosporin A specific antibody and cyclosporin A detection reagent |
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