CN101968492B - Particle-enhanced turbidimetric immune assay kit for detecting adiponectin and preparation method thereof - Google Patents
Particle-enhanced turbidimetric immune assay kit for detecting adiponectin and preparation method thereof Download PDFInfo
- Publication number
- CN101968492B CN101968492B CN 201010526950 CN201010526950A CN101968492B CN 101968492 B CN101968492 B CN 101968492B CN 201010526950 CN201010526950 CN 201010526950 CN 201010526950 A CN201010526950 A CN 201010526950A CN 101968492 B CN101968492 B CN 101968492B
- Authority
- CN
- China
- Prior art keywords
- adiponectin
- preparation
- mass percent
- latex
- particle
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Landscapes
- Peptides Or Proteins (AREA)
Abstract
The invention relates to a particle-enhanced turbidimetric immune assay kit for detecting adiponectin and a preparation method thereof, in particular to a detection method of a turbidimetric immune assay. The particle-enhanced turbidimetric immune assay kit for detecting the adiponectin comprises a kit body, an application liquid bottle and a latex suspension bottle coating an antiviral adiponectin monoclonal antibody, wherein the application liquid bottle is filled with application liquid; the application liquid comprises a surfactant, a preservative, a macromolecular accelerator, sodium chloride and a buffering agent; the latex suspension bottle coating the antiviral adiponectin monoclonal antibody is filled with a latex suspension coating the antiviral adiponectin monoclonal antibody; and the latex suspension coating the antiviral adiponectin monoclonal antibody comprises latex coating the antiviral adiponectin monoclonal antibody, the surfactant, a stabilizer and the buffering agent. The preparation method comprises the following steps of: preparing a latex antibody; preparing the latex suspension coating the antiviral adiponectin monoclonal antibody; preparing the application liquid; and finally preparing adiponectin series standard products.
Description
Technical field
The present invention relates to the turbid detection method of a kind of immune transmittance, especially relate to a kind of particle-enhanced turbidimetric immunoassay kit that detects adiponectin and preparation method thereof.
Background technology
Adiponectin (adiponectin) also claim Acrp30, GBP28 or AdipoQ, to secrete a kind of neurophysin by adipocyte, can be in human plasma stable existence, concentration range is about 3.0～17.0mg/L, account for 0.01% ([1] Arita Y of plasma proteins, Kihara S, Ouchi N, et al.Paradoxical decrease of an adipose-specific protein, adiponectin, inobesity[J] .Biochem Biophys Res Commun, 1999,257 (1): 79-83).Along with deepening continuously to adiponectin research, find, the concentration of adiponectin in obesity patient and type ii diabetes patients serum is starkly lower than non-overweight people, the reduction of its concentration has caused insulin resistance and hyperinsulinemia, and adiponectin has the effect of atherosclerosis Mottling formation, therefore, the genesis of adiponectin and obesity patient's type ii diabetes and coronary heart disease is closely related, the obvious reduction that is adiponectin is indicating that the generation of type ii diabetes and coronary heart disease obviously increases ([2] Zhang MH, Spies C, Ali S, et al.Adiponectinand inducible ischemia in patients with stable coronary heart disease:data from the Heart and Soulstudy[J] .Atherosclerosis, 2009,205 (1): 233-238; [3] Kawano J, Arora R.The role of adiponectin inobesity, diabetes, and cardiovascular disease[J] .J Cardiometab Syndr.2009,4 (1): 44-49).Measure adiponectin in the blood, significant to the diagnosis and prognosis of these diseases.
At present, the clinical diagnosis technology of adiponectin mainly comprises: radioimmunology (RIA) ([4] Schulze MB, Shai I, RimmEB, et al.Adiponectin and future coronary heart disease events among men with type 2diabetes[J] .Diabetes, 2005,54 (2): 534-539), enzyme linked immune assay (ELISA) ([5] Urbonaviciene G, Frystyk J, Flyvbjerg A, et al.Association of serum adiponectin with risk for cardiovascular events in patientswith peripheral arterial disease[J] .Atherosclerosis, 2010,210 (2): the method such as 619-624), their advantage is that sensitivity is higher, shortcoming is complex operation, length consuming time is subject to manual operation and extraneous factor and disturbs, and automaticity is low, and radioactively labelled substance can produce harm to the operator, to environment.These drawbacks limit applying of existing adiponectin detection method, make it can't be widely used in clinical diagnosis and research work.
Particle-enhanced turbidimetric immune assay method (particle-enhanced turbidimetric immunoassay, PETIA) be occur in recent years a kind of comparatively stable, body fluid albumen homogeneous phase immunoturbidimetry detection method accurately, its ultimate principle is surface-crosslinked monoclonal or the polyclonal antibody of certain grain size latex particle, the microballoon of antibody is arranged after antigen is combined when crosslinked, can flock together rapidly at short notice, change the absorbance of reactant liquor.And the concentration of this change and tested antigen has stronger correlativity, can reflect within the specific limits the concentration of tested antigen.The mode that this technology strengthens by particle, amplified antigen-antibody reaction, remedied the not high deficiency of common turbidimetry sensitivity, turbidimetry good stability, advantage have easily and fast been inherited again simultaneously, overcome the shortcoming of ELISA and RIA method, more and more be widely used in the clinically quantitative detection of various traces of albumin.At present, Latex-enhanced immunoturbidimetric assay has been widely used in bladder chalone C ([6] Al-Turkmani MR, Law T, Kellogg MD.Performance evaluation of a particle-enhanced turbidimetric cystatin C assay on theHitachi 917analyzer[J] .Clin Chim Acta.2008,398 (1-2): 75-77), lipoprotein (a), c reactive protein, hs-CRP, the detection of antistreptolysin O (ASO), obtain good social benefit, come out but there is not yet so far this type of adiponectin detection kit both at home and abroad.
Summary of the invention
The object of the present invention is to provide a kind of particle-enhanced turbidimetric immunoassay kit that detects adiponectin and preparation method thereof.
The particle-enhanced turbidimetric immunoassay kit of detection adiponectin of the present invention, the latex suspension bottle that comprises box body, application liquid bottle and coated anti-human adiponectin monoclonal antibody, the latex suspension bottle of described application liquid bottle and coated anti-human adiponectin monoclonal antibody is positioned in the box body, dress is used liquid in the described application liquid bottle, and the composition of described application liquid and content by mass percentage thereof are surfactant 0.1%～0.5%, antiseptic 0.05%～1%, macromolecule accelerator 2%～8%, sodium chloride 0.1%～10% and buffering agent 50～200mmol/L; The latex suspension of the coated anti-human adiponectin monoclonal antibody of dress in the latex suspension bottle of described coated anti-human adiponectin monoclonal antibody, the composition of the latex suspension of described coated anti-human adiponectin monoclonal antibody and content by mass percentage thereof are coated anti-human adiponectin monoclonal antibody latex 0.5%～5%, surfactant 0.1%～0.5%, stabilizing agent 0.1%～0.5% and buffering agent 10～100mmol/L.
Surfactant in the described application liquid can be selected from Tween 20 or Triton-X100 etc.
Antiseptic in the described application liquid can be selected from Proclin 300 or Sodium azide etc.
Macromolecule accelerator in the described application liquid is optional from polyglycol 6000 (PEG6000) or PEG 8000 (PEG8000) etc.
Buffering agent in the described application liquid can be selected from trishydroxymethylaminomethane (Tris), phosphate or glycocoll etc.
Surfactant in the latex suspension of described coated anti-human adiponectin monoclonal antibody can be selected from Tween 20 or Triton-X100 etc.
Stabilizing agent in the latex suspension of described coated anti-human adiponectin monoclonal antibody can be selected from bovine serum albumin(BSA) (BSA) etc.
Buffering agent in the latex suspension of described coated anti-human adiponectin monoclonal antibody is selected from trishydroxymethylaminomethane (Tris), phosphate or glycocoll etc.
Described application liquid buffering agent and latex SB preferably are consistent.
The preparation method of the particle-enhanced turbidimetric immunoassay kit of described detection adiponectin may further comprise the steps:
1) preparation latex antibody
Two kinds of mouse anti human adiponectin monoclonal antibodies are micro-sphere crosslinked with aldehyde radical respectively, get two kinds of latex antibody suspensions;
In step 1) in, described that two kinds of mouse anti human adiponectin monoclonal antibodies are micro-sphere crosslinked with aldehyde radical respectively, can adopt following concrete grammar: in carbonate buffer solution with the aldehyde radical microballoon respectively with two kinds of mouse anti human adiponectin monoclonal antibody mixings, behind 37 ℃ of revolving reactions, add the NaBH of carbonate buffer solution preparation
4Solution behind 4 ℃ of revolving reactions, adds the glycine solution of carbonate buffer solution preparation again, again behind 4 ℃ of revolving reactions, stores liquid and cleans with cleaning, and gained latex antibody precipitation is resuspended in to clean and stores in the liquid, gets latex antibody suspension, places 4 ℃ of environment placements;
Described two kinds of mouse anti human adiponectin monoclonal antibodies can adopt clone:Adn37 antibody and clone:Adn26 antibody;
The aldehyde radical microballoon that described aldehyde radical microballoon can adopt Shanghai to provide according to the health bio tech ltd, the particle diameter of described aldehyde radical microballoon can be 100～300nm, is preferably 150nm, and the mass percent of aldehyde radical microballoon is 0.5%～5%;
Described carbonate buffer solution pH can be 8.5～9.6, is preferably 9.6; Mass percent is 20～100mmol/L, is preferably 50mmol/L.
The time of described 37 ℃ of revolving reactions can be 4～16h, is preferably 16h;
Described NaBH
4The mass percent of solution is 0.05～0.5g/L, is preferably 0.1g/L;
The time of described 4 ℃ of revolving reactions can be 2～4h, is preferably 4h;
The mass percent of described glycine solution can be 10～100mmol/L, is preferably 50mmol/L;
The described again time of 4 ℃ of revolving reactions can be 2～24h, is preferably 16h;
Described cleaning is stored liquid pH and be can be 7.4～8.8, is preferably 8.0, comprises buffering agent, surfactant and stabilizing agent.Buffering agent can be phosphate, also can be trishydroxymethylaminomethane (Tris) or glycocoll, buffering agent shared mass percent in cleaning storage liquid is 10～100mmol/L, be preferably 25mmol/L, the mass percent of surfactant and stabilizing agent is respectively 0.1%～0.5% and 0.05%～0.5%, is preferably 0.2% and 0.1%.
2) the latex suspension of the coated anti-human adiponectin monoclonal antibody of preparation
Respectively with step 1) two kinds of latex antibody suspensions of preparation mix, and must be coated with the latex suspension of anti-human adiponectin monoclonal antibody;
In step 2) in, described two kinds of latex antibody suspensions mix preferably two kinds of latex antibody suspension equal-volumes mixing.
3) Application and preparation liquid
Surfactant, antiseptic, macromolecule accelerator, sodium chloride and buffering agent are dissolved in the water in the lump fully, get application liquid;
In step 3) in, described buffering agent kind and step 2) in clean to store that buffering agent is consistent in the liquid; Described application liquid pH value can be 7.4～8.0, is preferably 8.0; The shared mass percent of described surfactant can be 0.1%～0.5%, be preferably 0.2%, the shared mass percent of antiseptic can be 0.05%～1%, is preferably 0.1%, the shared mass percent of macromolecule accelerator can be 2%～8%, is preferably 4%; The shared mass percent of sodium chloride can be 0.1%～10%, is preferably 1.2%, and the mass percent of buffering agent can be 50～200mmol/L, is preferably 100mmol/L.
4) preparation adiponectin series standard product
Select the adiponectin standard items, use calf serum to carry out serial dilution: S6:40 μ g/ml, S5:20 μ g/ml, S4:10 μ g/ml, S3:5 μ g/ml, S2:1 μ g/mlS1:0.2 μ g/mlS0: calf serum, packing gets final product.
In step 4) in, described adiponectin standard items can be selected commercialization high-purity adiponectin standard items; Described packing can be adopted the packing of 0.5ml/ bottle.
Below provide the particle-enhanced turbidimetric immunoassay kit that adopts described detection adiponectin and carry out the actual conditions that adiponectin detects at Biochemical Analyzer:
Setup parameter on Biochemical Analyzer:
Temperature of reaction: 37 ℃;
Analytical approach: 2 end-point methods;
Predominant wavelength: 570nm, commplementary wave length: 800nm (can);
The latex suspension (R2) of sample size/application liquid (R1)/coated anti-human adiponectin monoclonal antibody: 5 μ l/200 μ l/50 μ l;
The Direction of Reaction: rise.
The present invention adopts anti-human adiponectin monoclonal antibody to be coated with the aldehyde radical microballoon, mode by the particle enhancing, amplified antigen-antibody reaction, remedied the not high deficiency of common turbidimetry sensitivity, turbidimetry good stability, advantage have easily and fast been inherited again simultaneously, overcome ELISA and RIA method complicated operation, bad etc. the shortcoming of repeatability, realize Acrp30 quantitatively fast detecting human adiponectin level in enormous quantities on biochemical instruments, be summed up the outstanding advantages of the following aspects: (1) is simple to operate; (2) sensitivity is higher, can reach 0.2 μ g/ml; (3) be not subject to manual operation and extraneous factor and disturb, detect stability and good reproducibility, can reflect more truly the content of measured matter; (4) can utilize Biochemical Analyzer to detect, easily realize robotization being suitable for clinical application.
Description of drawings
Fig. 1 is that the structure of the particle-enhanced turbidimetric immunoassay kit embodiment of detection adiponectin of the present invention forms schematic diagram.
Embodiment
Referring to Fig. 1, the particle-enhanced turbidimetric immunoassay kit embodiment of detection adiponectin of the present invention comprises box body 1, use the latex suspension bottle 3 of liquid bottle 2 and coated anti-human adiponectin monoclonal antibody, the latex suspension bottle 3 of described application liquid bottle 2 and coated anti-human adiponectin monoclonal antibody is positioned in the box body 1, dress is used liquid in the described application liquid bottle, and the composition of described application liquid and content by mass percentage thereof are surfactant 0.1%～0.5%, antiseptic 0.05%～1%, macromolecule accelerator 2%～8%, sodium chloride 0.1%～10% and buffering agent 50～200mmol/L; The latex suspension of the coated anti-human adiponectin monoclonal antibody of dress in the latex suspension bottle of described coated anti-human adiponectin monoclonal antibody, the composition of the latex suspension of described coated anti-human adiponectin monoclonal antibody and content by mass percentage thereof are coated anti-human adiponectin monoclonal antibody latex 0.5%～5%, surfactant 0.1%～0.5%, stabilizing agent 0.1%～0.5% and buffering agent 10～100mmol/L.
Preparation detects the particle-enhanced turbidimetric immunoassay kit of adiponectin
Take 4mg aldehyde radical microballoon as example, the key step of the present embodiment comprises:
1. prepare latex antibody
Select two kinds of mouse anti human adiponectin monoclonal antibodies (clone:Adn37, clone:Adn26) of high-purity, high-affinity.Use 20mmol/L carbonate buffer solution (pH9.6) with the aldehyde radical microballoon respectively with two kinds of antibody with 10: 1 quality than mixing, 37 ℃, revolving reaction 16h, this moment, total reaction volume was 400 μ l; After reaction finishes, add the 4mg/mlNaBH of above-mentioned carbonate buffer solution preparation
4Solution 20 μ l, 4 ℃, reaction 4h; Add again closed reagent 1mol/L glycine solution 20 μ l, 4 ℃, reaction 16h; (contain Tween 200.2%, BSA 0.1%, pH7.4) cleans 3 times for 25mmol/L TBS damping fluid; Precipitation is resuspended in 4ml25mmol/L TBS damping fluid, and (contain Tween 20 0.2%, BSA 0.1%, pH7.4), latex suspension concentration is 1mg/ml.
2. two kinds of latex antibody mix, the latex suspension of the coated anti-human adiponectin monoclonal antibody of preparation
Two kinds of latex antibody suspension equal-volumes that will prepare respectively mix, and this is R2 latex suspension.
3. Application and preparation liquid
The 50mmol/Ltris damping fluid of pH8.0 includes 12% NaCl, 4% PEG6000, and 0.1%Proclin 300.
4. the preparation of adiponectin series standard product
Select commercialization high-purity adiponectin standard items, use calf serum to carry out serial dilution: S6:40 μ g/ml, S5:20 μ g/ml, S4:10 μ g/ml, S3:5 μ g/ml, S2:1 μ g/ml, S1:0.2 μ g/mlS0: calf serum, the packing of 0.5ml/ bottle gets final product.
Preparation detects the particle-enhanced turbidimetric immunoassay kit of adiponectin
Take 4mg aldehyde radical microballoon as example, the key step of the present embodiment comprises:
1. prepare latex antibody
With the aldehyde radical microballoon respectively with two kinds of antibody with 20: 1 quality than mixing, remaining with embodiment 1.
2. two kinds of latex antibody mix, the latex suspension of the coated anti-human adiponectin monoclonal antibody of preparation
With embodiment 1.
3. Application and preparation liquid
The 50mmol/Ltris damping fluid of pH7.4 includes 12% NaCl, 6% PEG6000, and 0.1%Proclin 300.
4. the preparation of adiponectin series standard product
With embodiment 1.
Preparation detects the particle-enhanced turbidimetric immunoassay kit of adiponectin
1. prepare latex antibody
To add 4mg/ml NaBH in the step 1 preparation latex antibody
4The amount of solution changes 10 μ l into, and 4 ℃, the reaction time changes 2h into, and is remaining with embodiment 1.
2. two kinds of latex antibody mix, the latex suspension of the coated anti-human adiponectin monoclonal antibody of preparation
With embodiment 1.
3. Application and preparation liquid
The 200mmol/Ltris damping fluid of pH8.0 includes 5.0% NaCl, 6% PEG8000, and 0.1%Proclin 300.
4. the preparation of adiponectin series standard product
With embodiment 1.
Embodiment 4
Preparation detects the particle-enhanced turbidimetric immunoassay kit of adiponectin
1. prepare latex antibody
With embodiment 1 step 1) amount that adds closed reagent 1mol/L glycine solution in the preparation latex antibody changes 10 μ l into, and 4 ℃, the reaction time changes 4h into, Tween 20 changes 0.5% in the 25mmol/L TBS damping fluid, BSA changes 0.5%, pH into and changes 8.0 into, and is remaining with embodiment 1.
2. two kinds of latex antibody mix, the latex suspension of the coated anti-human adiponectin monoclonal antibody of preparation
With embodiment 1.
3. Application and preparation liquid
With embodiment 1.
4. the preparation of adiponectin series standard product
With embodiment 1.
Embodiment 5
Carry out the detection of adiponectin on the Biochemical Analyzer
Setup parameter on 7060 type Biochemical Analyzers: 37 ℃ of temperature of reaction; Analytical approach: 2 end-point methods; Predominant wavelength: 570nm, commplementary wave length: 800nm; Sample size/R1/R2:5 μ l/250 μ l/50 μ l; The Direction of Reaction: rise.Read point is respectively at 20 and 34 points.After selecting the multiple spot calibration, instrument is finished calibration automatically, and behind the calibration OK, the adiponectin that carries out conventional serum specimen detects, and the result is through the automatic directly report numerical value take mg/L as unit that converts of instrument.
Claims (12)
1. detect the particle-enhanced turbidimetric immunoassay kit of adiponectin, it is characterized in that comprising the latex suspension bottle of box body, application liquid bottle and coated anti-human adiponectin monoclonal antibody, the latex suspension bottle of described application liquid bottle and coated anti-human adiponectin monoclonal antibody is positioned in the box body, dress is used liquid in the described application liquid bottle, and the composition of described application liquid and content by mass percentage thereof are surfactant 0.1%～0.5%, antiseptic 0.05%～1%, macromolecule accelerator 2%～8%, sodium chloride 0.1%～10% and buffering agent 50～200mmol/L; The latex suspension of the coated anti-human adiponectin monoclonal antibody of dress in the latex suspension bottle of described coated anti-human adiponectin monoclonal antibody, the composition of the latex suspension of described coated anti-human adiponectin monoclonal antibody and content by mass percentage thereof are coated anti-human adiponectin monoclonal antibody latex 0.5%～5%, surfactant 0.1%～0.5%, stabilizing agent 0.1%～0.5% and buffering agent 10～100mmol/L.
2. the particle-enhanced turbidimetric immunoassay kit of detection adiponectin as claimed in claim 1 is characterized in that the surfactant in the described application liquid is selected from Tween 20 or Triton-X100; Antiseptic in the described application liquid is selected from Proclin 300 or Sodium azide; Macromolecule accelerator in the described application liquid is selected from Macrogol 6000 or PEG 8000; Buffering agent in the described application liquid is selected from trishydroxymethylaminomethane, phosphate or glycocoll.
3. the particle-enhanced turbidimetric immunoassay kit of detection adiponectin as claimed in claim 1 is characterized in that the surfactant in the latex suspension of described coated anti-human adiponectin monoclonal antibody is selected from Tween 20 or Triton-X100; Stabilizing agent in the latex suspension of described coated anti-human adiponectin monoclonal antibody is selected from bovine serum albumin(BSA); Buffering agent in the latex suspension of described coated anti-human adiponectin monoclonal antibody is selected from trishydroxymethylaminomethane, phosphate or glycocoll.
4. the preparation method of the particle-enhanced turbidimetric immunoassay kit of detection adiponectin as claimed in claim 1 is characterized in that may further comprise the steps:
1) preparation latex antibody suspension
Two kinds of mouse anti human adiponectin monoclonal antibodies are micro-sphere crosslinked with aldehyde radical respectively, get two kinds of latex antibody suspensions;
Described that two kinds of mouse anti human adiponectin monoclonal antibodies are micro-sphere crosslinked with aldehyde radical respectively, adopt following concrete grammar: in carbonate buffer solution with the aldehyde radical microballoon respectively with two kinds of mouse anti human adiponectin monoclonal antibody mixings, behind 37 ℃ of revolving reactions, add the NaBH of carbonate buffer solution preparation
4Solution behind 4 ℃ of revolving reactions, adds the glycine solution of carbonate buffer solution preparation again, again behind 4 ℃ of revolving reactions, stores liquid and cleans with cleaning, and gained latex antibody precipitation is resuspended in to clean and stores in the liquid, gets latex antibody suspension, places 4 ℃ of environment placements;
Described two kinds of mouse anti human adiponectin monoclonal antibodies adopt clone:Adn37 antibody and clone:Adn26 antibody; The particle diameter of described aldehyde radical microballoon is 100～300nm, and the mass percent of aldehyde radical microballoon is 0.5%～5%;
2) the latex suspension of the coated anti-human adiponectin monoclonal antibody of preparation
Two kinds of latex antibody suspensions with the step 1) preparation mix respectively, must be coated with the latex suspension of anti-human adiponectin monoclonal antibody;
3) Application and preparation liquid
Surfactant, antiseptic, macromolecule accelerator, sodium chloride and buffering agent are dissolved in the water in the lump fully, get application liquid;
4) preparation adiponectin series standard product
Select the adiponectin standard items, use calf serum to carry out serial dilution: S6:40 μ g/ml, S5:20 μ g/ml, S4:10 μ g/ml, S3:5 μ g/ml, S2:1 μ g/mlS1:0.2 μ g/mlS0: calf serum, packing gets final product.
5. the preparation method of the particle-enhanced turbidimetric immunoassay kit of detection adiponectin as claimed in claim 4 is characterized in that in step 1), and the particle diameter of described aldehyde radical microballoon is 150nm.
6. the preparation method of the particle-enhanced turbidimetric immunoassay kit of detection adiponectin as claimed in claim 4 is characterized in that in step 1), and described carbonate buffer solution pH is 8.5～9.6; Mass percent is 20～100mmol/L;
The time of described 37 ℃ of revolving reactions is 4～16h;
Described NaBH
4The mass percent of solution is 0.05～0.5g/L;
The time of described 4 ℃ of revolving reactions is 2～4h;
The mass percent of described glycine solution is 10～100mmol/L;
The time of described again 4 ℃ of revolving reactions is 2～24h.
7. the preparation method of the particle-enhanced turbidimetric immunoassay kit of detection adiponectin as claimed in claim 6 is characterized in that described carbonate buffer solution pH is 9.6; Mass percent is 50mmol/L;
The time of described 37 ℃ of revolving reactions is 16h;
Described NaBH
4The mass percent of solution is 0.1g/L;
The time of described 4 ℃ of revolving reactions is 4h;
The mass percent of described glycine solution is 50mmol/L;
The time of described again 4 ℃ of revolving reactions is 16h.
8. the preparation method of the particle-enhanced turbidimetric immunoassay kit of detection adiponectin as claimed in claim 4 is characterized in that in step 1), and it is 7.4～8.8 that liquid pH is stored in described cleaning, comprises buffering agent, surfactant and stabilizing agent; Buffering agent is phosphate, trishydroxymethylaminomethane or glycocoll, and buffering agent shared mass percent in cleaning storage liquid is 10～100mmol/L, and the mass percent of surfactant and stabilizing agent is respectively 0.1%～0.5% and 0.05%～0.5%.
9. the preparation method of the particle-enhanced turbidimetric immunoassay kit of detection adiponectin as claimed in claim 8, it is characterized in that it is 8.0 that liquid pH is stored in described cleaning, buffering agent shared mass percent in cleaning storage liquid is 25mmol/L, and the mass percent of surfactant and stabilizing agent is respectively 0.2% and 0.1%.
10. the preparation method of the particle-enhanced turbidimetric immunoassay kit of detection adiponectin as claimed in claim 4 is characterized in that in step 2) in, it is that two kinds of latex antibody suspension equal-volumes mix that described two kinds of latex antibody suspensions mix.
11. the preparation method of the particle-enhanced turbidimetric immunoassay kit of detection adiponectin as claimed in claim 4 is characterized in that in step 3), described buffering agent kind and step 2) in clean to store that buffering agent is consistent in the liquid; Described application liquid pH value is 7.4～8.0; The shared mass percent of described surfactant is 0.1%～0.5%, and the shared mass percent of antiseptic is 0.05%～1%, and the shared mass percent of macromolecule accelerator is 2%～8%; The shared mass percent of sodium chloride is 0.1%～10%, and the mass percent of buffering agent is 50～200mmol/L.
12. the preparation method of the particle-enhanced turbidimetric immunoassay kit of detection adiponectin as claimed in claim 11 is characterized in that described application liquid pH value is 8.0; The shared mass percent of described surfactant is 0.2%, and the shared mass percent of antiseptic is 0.1%, and the shared mass percent of macromolecule accelerator is 4%; The shared mass percent of sodium chloride is 1.2%, and the mass percent of buffering agent is 100mmol/L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010526950 CN101968492B (en) | 2010-10-29 | 2010-10-29 | Particle-enhanced turbidimetric immune assay kit for detecting adiponectin and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010526950 CN101968492B (en) | 2010-10-29 | 2010-10-29 | Particle-enhanced turbidimetric immune assay kit for detecting adiponectin and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101968492A CN101968492A (en) | 2011-02-09 |
| CN101968492B true CN101968492B (en) | 2013-01-09 |
Family
ID=43547677
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201010526950 Active CN101968492B (en) | 2010-10-29 | 2010-10-29 | Particle-enhanced turbidimetric immune assay kit for detecting adiponectin and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101968492B (en) |
Families Citing this family (21)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102253217B (en) * | 2011-04-07 | 2013-08-07 | 武汉生之源生物科技有限公司 | Detection kit of latex particle enhanced neutrophil gelatinase-associated lipid transfer protein |
| CN102331494B (en) * | 2011-06-16 | 2014-03-26 | 广州艺佳生物科技有限公司 | Sealing and stabilizing agent for microporous board |
| CN102507926B (en) * | 2011-10-12 | 2014-02-05 | 安徽信灵检验医学科技有限公司 | Gold-coated polystyrene nanoparticles and preparation method |
| CN102628868B (en) * | 2011-12-30 | 2014-09-17 | 北京九强生物技术股份有限公司 | Latex enhanced immunoturbidimetry kit for detection of asymmetric dimethylarginine content |
| CN102636653A (en) * | 2012-04-19 | 2012-08-15 | 上海蓝怡科技有限公司 | Compounded latex particle-enveloped cystatin C detection kit |
| CN102662064A (en) * | 2012-04-26 | 2012-09-12 | 苏州照康生物技术有限公司 | Immunonephelometry kit for detecting lipid carrier protein related to neutrophils gelatinase and preparation method thereof |
| CN102662059B (en) * | 2012-05-11 | 2014-08-13 | 北京美康生物技术研究中心 | Latex-enhanced immunoturbidimetry kit for measuring Helicobacter pylori antibody as well as preparation method and application thereof |
| CN102879569A (en) * | 2012-09-18 | 2013-01-16 | 苏州照康生物技术有限公司 | Immunoturbidimetry kit for detecting cysteine proteinase inhibitor C and preparation method thereof |
| CN104237522B (en) * | 2012-12-03 | 2016-12-21 | 武汉生之源生物科技股份有限公司 | Adiponectin reagent box for detecting content and preparation method thereof |
| WO2014189624A1 (en) * | 2013-05-20 | 2014-11-27 | Silicon Biodevices, Inc. | Digital control of on-chip magnetic particle assay |
| CN103777023B (en) * | 2013-12-16 | 2015-04-08 | 深圳市安之酶生物技术有限公司 | Kit for measuring adiponectin by nano latex enhanced immune turbidimetry |
| CN103675299A (en) * | 2013-12-24 | 2014-03-26 | 上海北加生化试剂有限公司 | Kit and method for detecting concentration of fibronectin in urine |
| CN104407154B (en) * | 2014-12-05 | 2016-03-30 | 重庆中元生物技术有限公司 | A kind of high sensitivity latex enhancing immune turbidimetry kit |
| CN105759061B (en) * | 2016-02-26 | 2018-06-19 | 厦门大学附属中山医院 | Human serum adiponectin detection kit based on microparticle chemiluminescence immunoassay |
| CN106290822B (en) * | 2016-07-28 | 2018-06-19 | 武汉景川诊断技术股份有限公司 | Latex microsphere preparation method and application is immunized in d-dimer |
| CN110007091A (en) * | 2019-04-02 | 2019-07-12 | 杭州博谱医药科技有限公司 | A kind of adiponectin detection kit and preparation method thereof |
| CN110907639A (en) * | 2019-12-05 | 2020-03-24 | 四川新健康成生物股份有限公司 | Serum amyloid protein A detection kit and preparation method thereof |
| CN111239421A (en) * | 2020-02-19 | 2020-06-05 | 安徽大千生物工程有限公司 | Latex-enhanced immunoturbidimetry kit for quantitatively detecting adiponectin ADPN and preparation and application methods thereof |
| CN114112957B (en) * | 2021-11-26 | 2022-06-21 | 浙江夸克生物科技有限公司 | Adiponectin determination kit and application thereof |
| CN114137229A (en) * | 2021-12-03 | 2022-03-04 | 苏州普瑞斯生物科技有限公司 | Adiponectin detection reagent production process adopting latex enhanced immunoturbidimetry |
| CN114295840B (en) * | 2021-12-29 | 2023-04-07 | 中元汇吉生物技术股份有限公司 | Kit for high-sensitivity quantitative determination of adiponectin and preparation method thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN202066863U (en) * | 2010-10-29 | 2011-12-07 | 厦门大学附属中山医院 | Particle-enhanced turbidimetric immunoassay kit for adiponectin detection |
-
2010
- 2010-10-29 CN CN 201010526950 patent/CN101968492B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN202066863U (en) * | 2010-10-29 | 2011-12-07 | 厦门大学附属中山医院 | Particle-enhanced turbidimetric immunoassay kit for adiponectin detection |
Non-Patent Citations (6)
| Title |
|---|
| 人血清脂联素定量ELISA检测方法的建立及初步临床应用;李登清、陈琳磊等;《第二军医大学学报》;20090731;第30卷(第7期);850-853 * |
| 人血清脂联素酶联免疫吸附试验测定方法的建立及临床应用研究;魏文青、赵满仓等;《标记免疫分析与临床》;20090630;第16卷(第3期);166-169 * |
| 抗脂联素单克隆抗体的制备及免疫学特性分析;赵满仓、魏文青等;《华北国防医药》;20081231;第20卷(第6期);35-37 * |
| 李登清、陈琳磊等.人血清脂联素定量ELISA检测方法的建立及初步临床应用.《第二军医大学学报》.2009,第30卷(第7期),850-853. |
| 赵满仓、魏文青等.抗脂联素单克隆抗体的制备及免疫学特性分析.《华北国防医药》.2008,第20卷(第6期),35-37. |
| 魏文青、赵满仓等.人血清脂联素酶联免疫吸附试验测定方法的建立及临床应用研究.《标记免疫分析与临床》.2009,第16卷(第3期),166-169. |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101968492A (en) | 2011-02-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101968492B (en) | Particle-enhanced turbidimetric immune assay kit for detecting adiponectin and preparation method thereof | |
| CN202066863U (en) | Particle-enhanced turbidimetric immunoassay kit for adiponectin detection | |
| Eda et al. | Development of a new microparticle‐enhanced turbidimetric assay for C‐reactive protein with superior features in analytical sensitivity and dynamic range | |
| CN102749454B (en) | Full-scale C-reactive protein (CRP) colloidal gold immunoturbidimetric assay kit | |
| CN106324239B (en) | Latex microsphere preparation method and application is immunized in C reactive protein | |
| CN102253217B (en) | Detection kit of latex particle enhanced neutrophil gelatinase-associated lipid transfer protein | |
| CN101377492B (en) | Bladder chalone C determining reagent kit | |
| CN101452001B (en) | Quantitative determination RBP4 kit by chemiluminescence magnetic enzymoimmune method | |
| Ledue et al. | High sensitivity immunoassays for C-reactive protein: promises and pitfalls | |
| CN108303544A (en) | A kind of whole blood c reactive protein detection kit | |
| US20110318847A1 (en) | Cvd assay | |
| Nishimura et al. | Determination of adiponectin in serum using a latex particle-enhanced turbidimetric immunoassay with an automated analyzer | |
| CN102692504A (en) | D-dimer quantitative fluorescence immunoassay test strip and preparation method thereof | |
| Machida et al. | Determination of serum lipoprotein lipase using a latex particle-enhanced turbidimetric immunoassay with an automated analyzer | |
| CN102662064A (en) | Immunonephelometry kit for detecting lipid carrier protein related to neutrophils gelatinase and preparation method thereof | |
| CN106018299A (en) | Kit for measuring serum amyloid A and preparation method of kit | |
| Chen et al. | Chemiluminescence immunoassay for sensing lipoprotein-associated phospholipase A2 in cardiovascular risk evaluation | |
| CN108570104A (en) | Recombinating adiponectin antigen, antibody and adiponectin nano rubber latex enhances immunoturbidimetry kit | |
| CN106053862B (en) | Serum amyloid A protein（SAA）Determine kit | |
| Lachno et al. | Validation of assays for measurement of amyloid-β peptides in cerebrospinal fluid and plasma specimens from patients with Alzheimer's disease treated with solanezumab | |
| Bakker | Immunoturbidimetry of urinary albumin: prevention of adsorption of albumin; influence of other urinary constituents. | |
| WO2010013525A1 (en) | Method of assaying complex and kit to be used therefor | |
| CN110007091A (en) | A kind of adiponectin detection kit and preparation method thereof | |
| CN108918888A (en) | It is a kind of detect Endostatin latex enhancing immune than turbid kit and its application | |
| CN108362892A (en) | A kind of Procalcitonin colloid gold immune turbidimetry detection reagent |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20160816 Address after: Beijing road in Baohe District of Hefei city in Anhui province and the intersection of Dalian road 230051 Patentee after: ANHUI IPROCOM BIOTECHNOLOGY CO., LTD. Address before: Siming Lake Road Xiamen city Fujian province 361004 No. 201-209 Patentee before: Zhongshan Hospital Xiamen University |
|
| CP02 | Change in the address of a patent holder |
Address after: 230051 Floor 1-4 of Building 18, Liandong U Valley Phase I, southeast corner of busy road and Jilin Road intersection, Baohe Economic Development Zone, Hefei City, Anhui Province Patentee after: ANHUI IPROCOM BIOTECHNOLOGY CO., LTD. Address before: Beijing road in Baohe District of Hefei city in Anhui province and the intersection of Dalian road 230051 Patentee before: ANHUI IPROCOM BIOTECHNOLOGY CO., LTD. |
|
| CP02 | Change in the address of a patent holder | ||
| CP02 | Change in the address of a patent holder |
Address after: No.659 Lanzhou Road, Baohe District, Hefei City, Anhui Province 230051 Patentee after: ANHUI IPROCOM BIOTECHNOLOGY Co.,Ltd. Address before: 230051 Floor 1-4 of Building 18, Liandong U Valley Phase I, southeast corner of busy road and Jilin Road intersection, Baohe Economic Development Zone, Hefei City, Anhui Province Patentee before: ANHUI IPROCOM BIOTECHNOLOGY Co.,Ltd. |
|
| CP02 | Change in the address of a patent holder |