CN106434610B - Thrombin solution - Google Patents
Thrombin solution Download PDFInfo
- Publication number
- CN106434610B CN106434610B CN201610874963.3A CN201610874963A CN106434610B CN 106434610 B CN106434610 B CN 106434610B CN 201610874963 A CN201610874963 A CN 201610874963A CN 106434610 B CN106434610 B CN 106434610B
- Authority
- CN
- China
- Prior art keywords
- thrombin solution
- thrombin
- water
- sodium
- surfactant
- 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
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
- C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
- C12N9/6424—Serine endopeptidases (3.4.21)
- C12N9/6429—Thrombin (3.4.21.5)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
- C12Y304/21—Serine endopeptidases (3.4.21)
- C12Y304/21005—Thrombin (3.4.21.5)
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- 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
- G01N33/86—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood coagulating time or factors, or their receptors
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/745—Assays involving non-enzymic blood coagulation factors
- G01N2333/75—Fibrin; Fibrinogen
Abstract
The invention discloses the stabilizer of thrombin solution, thrombin solution and relevant detection reagent, prepare the method for stable thrombin solution and the purposes of the stabilizer of thrombin solution.
Description
Technical field
The present invention relates to enzyme industrial circle and field of immunodetection, relate in particular to thrombin solution stabilizer,
Thrombin solution and relevant detection reagent prepare the method for stable thrombin solution and the stabilizer of thrombin solution
Purposes.
Background technology
Fibrinogen (Fibrinogen, FIB) is the fibrinous precursor of the main protein as blood clot
Matter is generated by hepatic parenchymal cells.About 80% of fibrinogen in organism is distributed in blood plasma (in normal adult about
200~400mg/dL), remaining is distributed in tissue.Fibrinogen be containing through disulfide-bonded 3 pairs of polypeptides (A α, B β and
γ) the glycoprotein of chain, A α, B β chains form fibrin by the way that fibrin ferment is decomposed, and are generated to play thrombus, hemostasis blood
The main function of bolt.Clinically, increase in inflammation, reduced in hepatosis, DIC of height etc..
The clinical meaning of fibrinogen assay includes but not limited to：1. pathologic increases：(1) prethrombotic state and thrombus
Property disease when, body coagulation function enhancing, plasma fibrinogen increases, such as acute myocardial infarction, diabetes, the hypertension of pregnancy
Disease, atherosclerosis, malignant tumour etc..(2) albumen synthesis increases, such as connective tissue disease, Huppert's disease.(3) anti-
Answering property increases, such as after acute infection, acute nephritis, burn, shock, major operation.2. pathologic reduces：(1) consumption is excessive, leads
Plasma content is caused to reduce, such as DIC.(2) Fibrinolytic Activities enhance, and FIB is decomposed, such as primary hyperfibrinolysis disease.(3)
Synthesis is reduced, such as serious hepatitis, hepatic sclerosis.
The measuring method of fibrinogen includes fibrin ferment additive process (Clauss methods), Clotting-time method (PT algorithms), exempts from
Epidemic disease turbidimetry (TIA), emulsion technique etc. generally use fibrin ferment additive process.
Under fibrin ferment additive process, the content of fibrinogen ultimately forms fibre according to fibrinogen and thrombin action
The principle of fibrillarin measures, and standard curve is made by reference blood plasma of international standard substance, with fibrin ferment come when measuring the clotting of plasma
Between, i.e., thrombin time (Thrombin Time, TT), gained plasma coagulation time are in negative with fibrinogen concentration in blood plasma
Correlation, to obtain the content of fibrinogen.
Specifically, thrombin time refers to the time of the clotting of plasma after standardized factor is added in blood plasma.
Under thrombin action, the fibrinogen in blood plasma to be checked is changed into fibrin.When anticoagulant substances in blood plasma to be checked
When increasing, thrombin time extends.When blood plasma to be checked is in situations such as acid, test object is with abnormal fibrous proteinemia
Under, thrombin time shortens.
For example, thrombin time extension sees plasma fibrinogen attenuating or textural anomaly；Clinical application heparin, or
Heparin sample anticoagulant substances when hepatopathy, nephrosis and systemic loupus erythematosus increase；Fibrinolytic system function is hyperfunction.
The fibrin ferment used in fibrinogen assay, cuts peptide from the factor as its precursor and forms tool
There is α-fibrin ferment of coagulation activity.Known this α-fibrin ferment meeting selfdecomposition is solidifying at the β-without coagulation activity of low molecular weight
Hemase, and then resolve into γ-fibrin ferment when long-term preservation, is usually freeze-dried.
For measuring the reagent of fibrinogen (FIB) content or thrombin time (TT), existing reagent is
Freeze-dried reagent in use, is required to redissolve in addition to expensive, inconvenient to use, and will necessarily bring and redissolve
Volumetric errors are caused in journey, and result is caused deviation occur.And this problem is not present in liquid reagent, and it is easy to use, that is, it opens
It uses, difference between batch smaller is as a result more acurrate.Therefore urgently need to develop the stabilizer that can stablize thrombin solution and
Stable thrombin solution product.
Invention content
It is molten to improve fibrin ferment for the defect that the technical problem to be solved by the present invention is to be not easy to preserve for thrombin solution
The stability of liquid reduces to extend the holding time and preserves cost, easy to use, reduces experimental error.
Present inventors discovered unexpectedly that lactate ion has with glutamate ion stablizes thrombin solution
Synergistic effect.
The present invention provides the stabilizers for thrombin solution, and it includes water-soluble lactic acid compounds and water-soluble paddy ammonia
Acid compound.In one embodiment, the stabilizer is by water-soluble lactic acid compound and water-soluble glutamic acid compounds group
At.In another embodiment, the stabilizer includes water-soluble lactic acid compound, water-soluble glutamic acid compounds and molten
Agent.In another embodiment in addition, the stabilizer includes water-soluble lactic acid compound, water-soluble glutamic acid chemical combination
Object, solvent and those skilled in the art being capable of conventional use of auxiliary element (such as buffer, surfactant, preservatives
Deng).The solvent can be one in the solvent that water, organic solvent or those skilled in the art routinely can determine or use
Kind or two or more combinations.Preferably, the solvent is water.
Preferably, in the stabilizer for thrombin solution of the invention, water-soluble lactic acid compound and water-soluble paddy ammonia
The ratio of acid compound is 1:50 to 50:1.Preferably, the upper limit of the ratio can be 40:1、30:1、20:1、10:1、9:
1、8:1、7:1、6:1、5:1、4:1、3:1、2:1, the lower limit of the ratio can be 1:40、1:30、1:20、1:10、1:9、1:
8、1:7、1:6、1:5、1:4、1:3、1:2, the ratio can be above-mentioned upper limit value and the range that lower limiting value arbitrarily combines.It is more excellent
Selection of land, the ratio can be 1:1.
Preferably, the water-soluble lactic acid compound can be water-soluble lactic acid salt.The water-soluble lactic acid salt can be
The combination of one or more of calcium lactate, lithium lactate, magnesium lactate or zinc lactate.Preferably, the water-soluble lactic acid salt
It is calcium lactate, lithium lactate or combinations thereof.Most preferably, the water-soluble lactic acid salt is calcium lactate or lithium lactate.
Water-soluble lactic acid compound used in the present invention is commercially available, such as from Shanghai Aladdin biochemical technology stock
The trade mark of part Co., Ltd (Shanghai Jing Chun biochemical technologies limited liability company) be Aladdin (Aladdin) calcium lactate (for example,
Article No. C110506), from Sigma-Aldrich Co., Ltd (Sigma-Aldrich LLC) calcium lactate (for example,
Product identification 21175), the lithium lactate etc. from Sinopharm Chemical Reagent Co., Ltd..
Preferably, the water-soluble glutamic acid compounds can be water-soluble glutamate.It is highly preferred that the water solubility
Glutamate is sodium glutamate, potassium glutamate or combinations thereof.Most preferably, the water-soluble glutamate is sodium glutamate.
Water solubility glutamic acid compounds used in the present invention are commercially available, such as are had from Shanghai sky biological reagent
The sodium glutamate of limit company, the trade mark from the Shanghai bio tech ltd Yuan Ye are the potassium glutamate of source leaf (for example, article No.
S20427) etc..
The present invention also provides the thrombin solutions of the stabilizer comprising the present invention.
The thrombin solution that the stabilizer of the present invention is applicable in is (referred to as " thrombin solution of the invention ") to make fibrin ferment
The mixing of aqueous solution, organic solvent solution or water and organic solvent made of being suspended or dissolved in water and/or organic solvent
Solution, preferably aqueous solution or water and organic solvent mixed solution.The organic solvent, as long as point of fibrin ferment will not be caused
Solution and activity suppression etc., and its final use is not influenced, there is no particular limitation, such as dimethyl sulfoxide (DMSO), glycerine, poly- second
Glycol, polypropylene glycol etc..One or more of these substances can also be applied in combination.
Preferably, it in thrombin solution of the invention, is calculated with the total volume of thrombin solution, water-soluble lactic acid compound
Concentration be 1g/L to 20g/L；It is highly preferred that in the thrombin solution of the present invention, calculated with the total volume of thrombin solution, water
The concentration of dissolubility polylactides is 2g/L to 10g/L；It is highly preferred that in the thrombin solution of the present invention, with thrombin solution
Total volume calculate, the concentration of water-soluble lactic acid compound is 2g/L, 3g/L, 4g/L, 5g/L, 6g/L, 7g/L, 8g/L, 9g/L
Or 10g/L.It is further preferred that in the thrombin solution of the present invention, calculated with the total volume of thrombin solution, water-soluble lactic acid
The concentration of compound is 7.7g/L.
Preferably, it in thrombin solution of the invention, is calculated with the total volume of thrombin solution, water-soluble glutamic acid chemical combination
The concentration of object is 1g/L to 150g/L；It is highly preferred that in the thrombin solution of the present invention, with the total volume meter of thrombin solution
It calculates, the concentration of water-soluble glutamic acid compounds is 10g/L to 100g/L；It is highly preferred that in the thrombin solution of the present invention, with solidifying
The total volume of hemase solution calculates, the concentration of water-soluble glutamic acid compounds be 10g/L, 15g/L, 20g/L, 25g/L, 30g/L,
35g/L、40g/L、45g/L、50g/L、55g/L、60g/L、65g/L、70g/L、75g/L、80g/L、85g/L、90g/L、95g/L
Or 100g/L；It is further preferred that in the thrombin solution of the present invention, calculated with the total volume of thrombin solution, water-soluble paddy
The concentration of propylhomoserin compound is 50g/L.
The thrombin solution of the present invention can also include buffer, surfactant and preservative.
Preferably, the buffer can be 4- hydroxyethyl piperazineethanesulfonic acids (N- (2- ethoxys) piperazine-N'-2- ethane
Sulfonic acid；Molecular formula:C8H18N2O4S, also known as HEPES acid), citric acid, phosphoric acid, acetic acid, imidazoles, 3- (N- morpholines) third sulphur
(MOPS), two (2- ethoxys) imido grpup three (methylol) methane (BIS-TRIS), trishydroxymethylaminomethane (TRIS), 3-
(N- morpholinyls) -2- hydroxy-propanesulfonic acids (MOPSO), N- (2- acetylaminos)-imino-diacetic acetic acid (ADA) or 2-morpholine ethane sulfonic acid
One or more of (MES) combination；It is highly preferred that buffer is 4- hydroxyethyl piperazineethanesulfonic acids.
Buffer used in the present invention is commercially available, such as the 4- from Suzhou City Bake bio tech ltd
Hydroxyethyl piperazineethanesulfonic acid, 3- (N- morpholinyls) -2- hydroxyls third from uncommon love (Shanghai) the chemical conversion industry Development Co., Ltd of ladder
The 2- morpholines that sulfonic acid (for example, product coding H0671), the trade mark from Sa En chemical technologies (Shanghai) Co., Ltd. are An Naiji
Ethanesulfonic acid (for example, goods number D090002).
There is no particular limitation as long as the amount with buffer capacity for the additive amount of the buffer, people in the art
Member can routinely determine the content for the buffer for being suitble to use.But the inventors found that the fibrin ferment of the present invention is molten
It in liquid, is calculated with the total volume of thrombin solution, when the concentration of buffer is following values range, is more advantageous to and realizes this hair
Bright purpose：Preferably, it in thrombin solution of the invention, is calculated with the total volume of thrombin solution, the concentration of buffer is
1g/L to 50g/L；It is highly preferred that in the thrombin solution of the present invention, calculated with the total volume of thrombin solution, buffer it is dense
Degree is 5g/L to 24g/L；It is highly preferred that in the thrombin solution of the present invention, calculated with the total volume of thrombin solution, buffer
Concentration be 5.9g/L to 23.8g/L；It is highly preferred that in the thrombin solution of the present invention, with the total volume meter of thrombin solution
Calculate, the concentration of buffer be 5.9g/L, 6g/L, 7g/L, 8g/L, 9g/L, 10g/L, 11g/L, 11.9g/L, 12g/L, 13g/L,
14g/L, 15g/L, 16g/L, 17g/L, 17.9g/L, 18g/L, 19g/L, 20g/L, 21g/L, 22g/L, 23g/L or 23.8g/L；
It is further preferred that in the thrombin solution of the present invention, calculated with the total volume of thrombin solution, the concentration of buffer is
11.9g/L。
The surfactant can be anionic surfactant, cationic surfactant, amphoteric ion table
The combination of one or more of face activating agent or nonionic surfactant.
The anionic surfactant can be lauryl sodium sulfate, dodecyl sodium sulfate, dodecyl-N-
The combination of one or more of sodium sarcosinate, sodium taurocholate, NaTDC or sodium taurodeoxycholate.
The cationic surfactant can be cetyl trimethylammonium bromide, four decyl ammonium bromides or dodecane
The combination of one or more of pyridinum chloride.
The zwitterionic surfactant can be 3- [(3- courages amido propyl) dimethyl ammonium] -1- propane sulfonic acid salt, 3-
[(3- courages amido propyl) dimethyl ammonium] -2- hydroxyl -1- propane sulfonic acid salt, palmityl lysolecithin, dodecyl-N- glycine betaines
Or the combination of one or more of dodecyl-Beta-alanine.
The nonionic surfactant can be octyl glucoside, heptyl glucosinolate, capryl-N- methyl Portugal
Osamine, polyoxyethylene lauryl ether, seven methylhexyl ether of polyoxyethylene, Triton X100, polyoxyethylene nonyl
One or both of base phenyl ether, polyoxyethylene fatty acid ester, sucrose fatty ester or polyoxyethylene sorbitol ester with
On combination.
Preferably, the surfactant is nonionic surfactant；It is highly preferred that the surfactant is poly-
Ethylene oxide Sorbitan Monooleate (also known as Tween 80).
Surfactant used in the present invention is commercially available, such as the tween from Chengdu Ke Long chemical reagents factory
80, come from Shanghai Aladdin biochemical technology limited liability company (Shanghai Jing Chun biochemical technologies limited liability company) trade mark be Ah
The Tween 80 (for example, article No. T104866) of Latin (Aladdin), the trade mark of the Shanghai bio tech ltd Yuan Ye are source leaf
Sucrose fatty ester (for example, article No. S30894) etc..
As long as the additive amount of the surfactant makes the stability-enhanced amount of thrombin solution, without special
It limits, those skilled in the art can routinely determine the content for the surfactant for being suitble to use.But the present inventor
It was found that in the thrombin solution of the present invention, calculated with the total volume of thrombin solution, when the concentration of surfactant is following number
When being worth range, it is more advantageous to and achieves the object of the present invention：Preferably, it is calculated with the total volume of thrombin solution, surfactant
Concentration be 10 μ l/L to 1000 μ l/L；It is highly preferred that being calculated with the total volume of thrombin solution, the concentration of surfactant is
50 μ l/L to 500 μ l/L；It is highly preferred that calculated with the total volume of thrombin solution, the concentration of surfactant be 100 μ l/L extremely
400μl/L；It is further preferred that being calculated with the total volume of thrombin solution, the concentration of surfactant is 100 μ l/L, 200 μ
L/L, 300 μ l/L or 400 μ l/L；It is further preferred that being calculated with the total volume of thrombin solution, the concentration of surfactant
It is 200 μ l/L.
The preservative can be Sodium azide (NaN3), Ciprofloxacin, one or both of propionic acid or sodium benzoate with
On combination.Preferably, the preservative is Sodium azide.
Preservative used in the present invention is commercially available, such as comes precious biotechnology (Shanghai) limited liability company westerly
Trade mark be Amresco Sodium azide (for example, article No. be 0639), come from Chengdu Hua Xia chemical reagent Co., Ltd Sodium azide
(the brilliant pure biochemical technology share in Shanghai is limited for (for example, article No. is H1100046), Shanghai Aladdin biochemical technology limited liability company
Company) trade mark be Aladdin (Aladdin) sodium benzoate (for example, article No. is S104128), come from Sigma-Aldrich
The preservative that the trade mark of Co., Ltd (Sigma-Aldrich LLC) is Proclin300 is (for example, product identification is
48914-U) etc..
There is no particular limitation as long as in prescribed limit for the additive amount of the preservative.Those skilled in the art can be with
The conventional content for determining the preservative for being suitble to use.But the inventors found that the present invention thrombin solution in, with
The total volume of thrombin solution calculates, and when the concentration of preservative is following values range, is more advantageous to the mesh for realizing the present invention
's：Preferably, it is calculated with the total volume of thrombin solution, the concentration of preservative is 0.1g/L to 5.0g/L；It is highly preferred that with solidifying
The total volume of hemase solution calculates, and the concentration of preservative is 0.2g/L to 2.5g/L；It is highly preferred that with the totality of thrombin solution
Product calculates, and the concentration of preservative is 0.4g/L to 1.3g/L；It is further preferred that being calculated with the total volume of thrombin solution, prevent
The concentration of rotten agent be 0.4g/L, 0.5g/L, 0.6g/L, 0.7g/L, 0.8g/L, 0.9g/L, 1.0g/L, 1.1g/L, 1.2g/L or
1.3g/L；It is further preferred that being calculated with the total volume of thrombin solution, the concentration of preservative is 1.0g/L.
The fibrin ferment used in the present invention can be the fibrin ferment of the animal origins such as people, pig, ox, by genetic engineering legal system
Standby fibrin ferment or commercially available drug.Fibrin ferment used in the present invention is commercially available, e.g. comes from one lattice pharmacy of Hunan
The fibrin ferment freeze-dried powder of Co., Ltd, coagulating from Sigma-Aldrich Co., Ltd (Sigma-Aldrich LLC)
Hemase preparation (for example, product identification is 10602400001) etc..
The additive amount of fibrin ferment in the thrombin solution of the present invention is as long as being suitble to the purpose of the present invention without special
It limits.It was found by the inventors of the present invention that in the thrombin solution of the present invention, calculated with the total volume of thrombin solution, fibrin ferment
Concentration can be 1KU/L to 120KU/L；It is highly preferred that in the thrombin solution of the present invention, with the total volume of thrombin solution
Calculate, the concentration of fibrin ferment can be 1KU/L, 2KU/L, 3KU/L, 4KU/L, 5KU/L, 6KU/L, 7KU/L, 8KU/L, 9KU/L,
10KU/L、20KU/L、30KU/L、40KU/L、50KU/L、60KU/L、70KU/L、80KU/L、90KU/L、100KU/L、110KU/
L or 120KU/L；It is highly preferred that in the thrombin solution of the present invention, calculated with the total volume of thrombin solution, fibrin ferment it is dense
Degree can be 2KU/L to 110KU/L, 5KU/L to 100KU/L, 10KU/L to 90KU/L, 15KU/L to 80KU/L, 20KU/L extremely
70KU/L, 25KU/L are to 60KU/L, 30KU/L to 50KU/L；It is further preferred that in the thrombin solution of the present invention, with blood coagulation
The total volume of enzyme solutions calculates, and the concentration of fibrin ferment can be 2KU/L, 4KU/L, 30KU/L, 70KU/L, 120KU/L.
Preferably, thrombin solution of the invention is calculated with the total volume of thrombin solution, including concentration is 7.7g/L
It is 4- hydroxyethyl piperazineethanesulfonic acids, the concentration of 11.9g/L is 1.0g/L that calcium lactate, concentration, which are the sodium glutamate of 50g/L, concentration,
Sodium azide, concentration are the polyoxyethylene sorbitan monooleates of 200 μ l/L.It is highly preferred that in above-mentioned thrombin solution, packet
It is the fibrin ferment of 2KU/L, 4KU/L, 30KU/L or 70KU/L containing concentration.
Preferably, thrombin solution of the invention is calculated with the total volume of thrombin solution, including concentration is 7.7g/L
It is 4- hydroxyethyl piperazineethanesulfonic acids, the concentration of 11.9g/L is 1.0g/L that lithium lactate, concentration, which are the sodium glutamate of 50g/L, concentration,
Sodium azide, concentration are the polyoxyethylene sorbitan monooleates of 200 μ l/L.It is highly preferred that in above-mentioned thrombin solution, packet
It is the fibrin ferment of 2KU/L, 4KU/L, 30KU/L or 70KU/L containing concentration.
The present invention also provides the methods for preparing thrombin solution, and the method includes using the step of the stabilizer of the present invention
Suddenly.
The present invention also provides the purposes that the stabilizer of the present invention is used to improve thrombin solution stability.
The present invention also provides the purposes that the thrombin solution of the stabilizer of the present invention or the present invention is used to prepare reagent.
The present invention also provides a kind of reagents, contain the stabilizer of the present invention or the thrombin solution of the present invention.
The reagent prepared using the stabilizer of the present invention or the thrombin solution of the present invention or the stabilization containing the present invention
The reagent of the thrombin solution of agent or the present invention can be referred to as the reagent of the present invention.
Preferably, the reagent is the reagent for detecting fibrinogen or thrombin time.
Preferably for the present invention for detecting the reagent of fibrinogen, fibrin ferment in thrombin solution contains
Amount is 30KU/L or 70KU/L；For the present invention for detecting the reagent of thrombin time, the fibrin ferment in thrombin solution
Content be 2KU/L or 4KU/L.
The present invention also provides a kind of kits, and it includes the reagents of the present invention.
The present inventors have noted that in the case where thrombin solution other components are constant, under different concentration of thrombin, contain
There is the measured value of the reagent of the thrombin solution of the present invention that can integrally improve, but this thrombin solution and correlation to the present invention
The implementation of the stability of product itself and relevant purposes, method does not have an impact, and the raising of measured value will not interfere this hair
The realization of the function of bright thrombin solution and Related product.In order to obtain the measured value in ideal range, people in the art
Member can realize above-mentioned target with the content of general adjustment fibrin ferment.
The thrombin solution of stabilizer containing the present invention is easy to use, without redissolving, avoids due to redissolving volume difference
Lead to error.Plug and play, difference between batch smaller are as a result more acurrate.
The thrombin solution stability prepared using the stabilizer of the present invention is strong, and 2-8 DEG C is stablized 1 year or more, 37 DEG C of stabilizations
20 days or more, 2-8 DEG C of corkage stability 10 days or more.Stabilizer of the invention can improve the stabilization of thrombin solution as a result,
Property, improve the storage form and condition of Related product, reduces the cost of production and application.
Specific implementation mode
The each component source used in the following embodiment of the present invention is respectively：Shanghai Aladdin biochemical technology share is limited
The trade mark of company (Shanghai Jing Chun biochemical technologies limited liability company) is the calcium lactate (article No. of Aladdin (Aladdin)
C110506), the lithium lactate from Sinopharm Chemical Reagent Co., Ltd., from the Shanghai bio tech ltd Yuan Ye
Trade mark is the potassium glutamate (for example, article No. S20427) of source leaf, the sodium glutamate of Shanghai sky biological reagent Co., Ltd, Suzhou
The 4- hydroxyethyl piperazineethanesulfonic acids (HEPES acid) of city Bake bio tech ltd, the tween of Chengdu Ke Long chemical reagents factory
80, the trade mark of precious biotechnology (Shanghai) limited liability company in west is the Sodium azide (article No. 0639) of Amresco, one lattice system of Hunan
The fibrin ferment freeze-dried powder of medicine Co., Ltd.
Embodiment 1
The preparation of thrombin time (TT) detection reagent of the stabilizer containing the present invention
HEPES acid 11.9g, calcium lactate 7.7g, sodium glutamate 50g, Sodium azide 1g, Tween 80 0.2g are weighed, distillation is added
It is dissolved in water, adjusts pH to 6.4 with sodium hydroxide, be settled to 1L.Fibrin ferment 4 (1000U/ branch) is taken, is buffered with above prepare
Liquid 1mL/ branch dissolves, and takes dissolving fibrin ferment 3.6mL (adjustable enzyme amount control difference between batch), is added in 1L buffer solutions and shakes up, TT inspections
Test agent, which is prepared, to be completed.
Product performance index
1. repeatability：The coefficient of variation (CV)≤5%；With normal Quality Control blood plasma retest 10 times, being averaged for measurement is calculated
ValueWith standard deviation (s).The coefficient of variation (CV) is calculated according to formula (1).Acquired results should meet wanting for following 3. stability
It asks.
In formula：
In S：Standard deviation
--- measure mean value
2. difference between batch：The coefficient of variation (CV)≤10%；Test the reagent of 3 different batches respectively with normal Quality Control blood plasma,
Each batch measures retest 10 times, calculates 30 measured value average valuesWith standard deviation (s), calculated according to formula (1)
The coefficient of variation (CV).
3. stability
2 DEG C~8 DEG C closed preservations of this product are newly opened one bottle of measurement Quality Control blood plasma, were compared with first day, relative deviation exists every time
In 15%, the term of validity 12 months.
Embodiment 2
The detection method of thrombin time (TT) detection reagent
100 μ L of blood plasma are taken, 37 DEG C preheat 2 minutes, 100 μ L of thrombin reagent are then added, in CA550 full automatic blood-coagulation instrument
(producer SYSMEX) is measured according to the specification of producer.
Points for attention：
1. blood sampling should avoid haemolysis, tissue fluid is prevented to be mixed into.Smooth when blood drawing, anti-freezing is abundant, can not there is sludged blood；
Blood platelet must be removed when separated plasma.
2. when sample collection anti-coagulants should not be made with EDTA and heparin.The ratio of blood and anti-coagulants should be accurate, blood sampling volume
Sample of the deviation more than 10% should not use.
3. if hematocrit≤20% or >=55%, need the ratio of adjustment blood sample and anti-coagulants, anti-coagulants ml
=0.00185 × blood ml × (100- hematocrits).
Measuring pipette and sample injector after 4. use is calibrated.
5. using the plastics of cleanliness without any pollution or the glassware of silication.
6. experimental temperature is controlled at 37 DEG C ± 1.0 DEG C.
7. reagent can change in proportion as needed with sample size.
8. reagent use finishes, bottleneck should be wiped clean, screws bottle cap as early as possible, puts back under the condition of storage of requirement and protects in time
It deposits.
Embodiment 3
The stability experiment of thrombin time (TT) detection reagent of the stabilizer containing the present invention
Thrombin time (TT) detection reagent is prepared according to the scheme of embodiment 1, uses CA550 full automatic blood-coagulation instrument (factory
Family SYSMEX) according to the stability of the reagent under the specification measurement the following conditions of producer
1. 37 DEG C accelerate the failure.
2. 2-8 DEG C of corkage stability.
3. 2-8 DEG C of long-time stability.
37 DEG C surely accelerate the failure, normal Quality Control blood plasma measured value, and measured value is stablized within January.It is shown in Table 1.
1 37 DEG C of table accelerates heat damage experiment
Stability experiment is opened, 2-8 DEG C is put in after corkage, 15 days, measured value was relatively stablized.It is shown in Table 2.
The 2-8 DEG C of corkage stability of table 2
2-8 DEG C of long-time stability, 15 months, measured value was relatively stablized.It is shown in Table 3.
3 long-time stability of table
Embodiment 4
The preparation of fibrinogen (FIB) detection reagent of the stabilizer containing the present invention
(1) reagent preparation：Weigh HEPES acid 11.9g, calcium lactate 7.7g, sodium glutamate 50g, Sodium azide 1g, Tween 80
0.2g is added in distilled water and dissolves, and adjusts pH to 6.4 with sodium hydroxide, is settled to 1L.Fibrin ferment 70 (1000U/ branch) is taken,
The dissolving of buffer solution 1mL/ branch is prepared with above, is all added in 1L buffer solutions and shakes up, FIB detection reagents, which are prepared, to be completed.
(2) prepared and diluted sample buffer：Weigh imidazoles 3.06g, sodium chloride 5.22g, sodium citrate 1.2g, Sodium azide
0.95g is added in distilled water and dissolves, and adjusts pH to 7.30 with sodium hydroxide, is settled to 1L.After stable in two days, pH is adjusted again
To 7.30.
Product performance index
1. accuracy：It takes reference material or correctness Quality Control object to test, in triplicate, obtains average value, counted according to formula (2)
Average value and reference material or the relative deviation of main calibration object are calculated, as a result should meet relative deviation should want in ± 15% range
It asks.Calculation formula：
In formula：
X-- detects mean value；
Xc-- reference materials or correctness Quality Control object sign value.
2. repeatability
Distinguish retest 10 times with normal Quality Control blood plasma and abnormal Quality Control blood plasma, and calculates the average value of measurementWith
Standard deviation (S).The coefficient of variation (CV) is calculated by formula (1), as a result should meet the requirement of the coefficient of variation (CV)≤8%.
3. difference between batch
The reagent of 3 different batches is tested with normal Quality Control blood plasma, each batch is tested 10 times, and it is flat to calculate 30 measured values
Mean valueWith standard deviation (S), the coefficient of variation (CV) is calculated according to formula (1), as a result should meet the coefficient of variation (CV)≤15%
Requirement.
4. stability
One bottle of measurement Quality Control blood plasma is newly opened in 2 DEG C~8 DEG C closed preservations of this product every time, and relative deviation is in 15%, the term of validity
12 months.
Embodiment 5
The detection method of fibrinogen (FIB) detection reagent
(1) standard curve is established
Reference blood plasma is diluted with dilution according to the form below, you can obtains series concentration：
FIB detection reagents pre-temperature is taken 3 minutes, until 37 DEG C.Take the reference blood plasma 100 μ l after dilution, 37 DEG C of pre-temperatures 2 minutes.
50 μ l pre-temperature reagents are added, at once mixing and timing, record setting time.It is with the obtained target level of reference blood plasma gradient dilution
Abscissa, corresponding setting time (second) is ordinate, and measurement result is mapped on double logarithmic curve, or input instrument is according to corresponding
Mathematical model automatically generate working curve.
(2) sample measures
Sample carries out 10 times of dilutions, preparation of the determination step with working curve with dilution buffer.It is automatically coagulated with CA550
Blood instrument (producer SYSMEX) is measured according to the specification of producer.
Points for attention：
1. blood sampling should avoid haemolysis, tissue fluid is prevented to be mixed into.Smooth when blood drawing, anti-freezing is abundant, can not there is sludged blood；
Blood platelet must be removed when separated plasma.
2. when sample collection anti-coagulants should not be made with EDTA and heparin.The ratio of blood and anti-coagulants should be accurate, blood sampling volume
Sample of the deviation more than 10% should not use.
3. if hematocrit≤20% or >=55%, need the ratio of adjustment blood sample and anti-coagulants, anti-coagulants ml
=0.00185 × blood ml × (100- hematocrits).
Measuring pipette and sample injector after 4. use is calibrated.
5. using the plastics of cleanliness without any pollution or the glassware of silication.
6. experimental temperature is controlled at 37 DEG C ± 1.0 DEG C.
7. reagent can change in proportion as needed with sample size.
8. reagent use finishes, bottleneck should be wiped clean, screws bottle cap as early as possible, puts back under the condition of storage of requirement and protects in time
It deposits.
Embodiment 6
The stability experiment of fibrinogen (FIB) detection reagent of the stabilizer containing the present invention
Fibrinogen (FIB) detection reagent is prepared according to the scheme of embodiment 4, uses CA550 full automatic blood-coagulation instrument (factory
Family SYSMEX) according to the stability of the reagent under the specification measurement the following conditions of producer.
1. 37 DEG C accelerate the failure.
2. 2-8 DEG C of corkage stability.
3. 2-8 DEG C of long-time stability.
37 DEG C accelerate the failure, normal Quality Control blood plasma measured value, and measured value is stablized within January.It is shown in Table 4.
4 37 DEG C of table accelerates heat damage experiment
Stability experiment is opened, 2-8 DEG C is put in after corkage, is opened 17 days, measured value is relatively stablized.It is shown in Table 5.
The 2-8 DEG C of corkage stability of table 5
2-8 DEG C of long-time stability, 416 days, measured value was relatively stablized.It is shown in Table 6.
The 2-8 DEG C of long-time stability of table 6
Embodiment 7
There is the synergistic effect for stablizing thrombin solution between water-soluble lactic acid compound and water-soluble glutamic acid compounds
Under conditions of its dependent variable is consistent, for only containing calcium lactate, only containing sodium glutamate and containing breast simultaneously
Three kinds of thrombin solutions of sour calcium and sodium glutamate measure the stabilization time at 37 DEG C, are nature controlling line with relative deviation 10%, see
Examine the testing result of above-mentioned thrombin solution.Testing result takes the mean value of 5 experiments.
The application also measures the stabilization time containing calcium lactate and potassium glutamate thrombin solution at 37 DEG C simultaneously, with
Relative deviation 10% is nature controlling line, observes the testing result of above-mentioned thrombin solution.Testing result takes the mean value of 3 experiments.
The content of each ingredient is respectively in thrombin solution in serial number 1-4 in table 7：Calcium lactate 7.7g/L (such as containing), paddy
Propylhomoserin sodium 50g/L (such as containing), potassium glutamate 50g/L (such as containing), HEPES acid 11.9g/L, Sodium azide 1g/L, Tween 80
200 μ l/L, fibrin ferment 4KU/L (solution of serial number 1-3) or 2KU/L (solution of serial number 4).
Table 7：Calcium lactate stablizes the synergistic effect of thrombin solution with water-soluble glutamic acid compounds
Table 8a calcium lactates stablize the experiment of thrombin solution with water-soluble glutamic acid compounds
From table 7, table 8a it is observed that when calcium lactate is existed simultaneously with water-soluble glutamic acid compounds, thrombin solution
Stabilization number of days longest.Therefore, exist between calcium lactate and water-soluble glutamic acid compounds stablize thrombin solution cooperate with effect
It answers.
The present invention also has chosen the thrombin solution containing lithium lactate and sodium glutamate further to verify water-soluble lactic acid
There is the synergistic effect for stablizing thrombin solution between compound and water-soluble glutamic acid compounds.When measuring the stabilization at 37 DEG C
Between, it is nature controlling line with relative deviation 10%, observes the testing result of above-mentioned thrombin solution.Testing result takes the equal of 3 experiments
Value.
Above-mentioned lithium lactate and sodium glutamate are respectively as the content of each ingredient in the thrombin solution of stabilizer：Lithium lactate
7.7g/L, sodium glutamate 50g/L, HEPES acid 11.9g/L, Sodium azide 1g/L, 200 μ l/L of Tween 80, fibrin ferment 30KU/L
(FIB reagents) or 2KU/L (TT reagents).The stability test of above-mentioned thrombin solution is as follows：
Table 8b lithium lactates stablize the experiment of thrombin solution with sodium glutamate
From table 8b as it can be seen that lithium lactate also has good synergisticing stable effect with sodium glutamate.
Therefore, it can be seen that between water-soluble lactic acid compound and water-soluble glutamic acid compounds and exist from above-mentioned experiment
Stablize the synergistic effect of thrombin solution.
Embodiment 8
Whether there is or not sodium glutamates to influence Experimental comparison to TT reagent stabilities.
According to the thrombin solution formula in embodiment 1, in the case of other components content is constant, measure at 37 DEG C respectively
The stability of thrombin solution containing sodium glutamate and without sodium glutamate.
Whether there is or not sodium glutamate influences for 9 37 DEG C of table
Formulation stability containing sodium glutamate and calcium lactate it can be seen from the test result of table 9 is substantially better than no paddy ammonia
The stability of the formula of sour sodium.
Embodiment 9
Stablizing effect of the amino acid or its salt of calcium lactate and other non-glutamic acids to thrombin solution
The content of each ingredient is respectively in thrombin solution：Calcium lactate 7.7g/L, glycine 50g/L (such as containing), smart ammonia
Sour 50g/L (such as containing), serine 50g/L (such as containing), glutamic acid 50g/L (such as containing), HEPES acid 11.9g/L, Sodium azide
1g/L, 200 μ l/L of Tween 80.Fibrin ferment is 4KU/L in TT reagents, and fibrin ferment is 70KU/L in FIB reagents.
It is nature controlling line with relative deviation 10%, according to table 10a in table 7 in embodiment 7 and the present embodiment, 11 experiment knot
Fruit, calcium lactate can not realize water-soluble glutamic acid compounds to thrombin solution with the amino acid of other non-glutamic acids or its salt
Stablizing effect.
Table 10a：Calcium lactate stablizes the synergistic effect of thrombin solution with the amino acid of other non-glutamic acids
Table 10b：Calcium lactate stablizes the synergistic effect of thrombin solution with glycine or valine
In embodiment 10-14, in thrombin solution when a certain component content variation, the content of other compositions follows following number
Value：Calcium lactate 7.7g/L, sodium glutamate 50g/L, HEPES acid 11.9g/L, Sodium azide 1g/L, 200 μ l/L of Tween 80.About solidifying
The content of enzyme in hemase solution, in TT reagents, the content of enzyme is 4KU/L；In FIB reagents, the content of enzyme is 70KU/L.
Embodiment 10
The content range of HEPES acid in thrombin solution
Be nature controlling line with 15% according to table 11, in thrombin solution the content range of HEPES acid can be 5.9g/L extremely
23.8g/L。
The content range of HEPES acid in 11 thrombin solution of table
Embodiment 11
The content range of calcium lactate in thrombin solution
It is nature controlling line with 15% according to table 12, the content range of calcium lactate can be 2g/L to 10g/ in thrombin solution
L。
The content range of calcium lactate in 12 thrombin solution of table
Embodiment 12
The content range of thrombin solution Glutamic Acid sodium
Be nature controlling line with 15% according to table 13, the content range of thrombin solution Glutamic Acid sodium can be 10g/L extremely
100g/L。
The content range of 13 thrombin solution Glutamic Acid sodium of table
Embodiment 13
The content range of Sodium azide in thrombin solution
Be nature controlling line with 15% according to table 14, in thrombin solution the content range of Sodium azide can be 0.4g/L extremely
1.3g/L。
The content range of Sodium azide in 14 thrombin solution of table
Embodiment 14
The content range of Tween 80 in thrombin solution
Be nature controlling line with 15% according to table 15, in thrombin solution the content range of Tween 80 can be 100 μ l/L extremely
400μl/L。
The content range of Tween 80 in 15 thrombin solution of table
Claims (12)
1. a kind of thrombin solution, it includes stabilizers, wherein the stabilizer includes water-soluble lactic acid salt, water-soluble glutamic acid
Salt, buffer, surfactant and preservative, wherein the water-soluble lactic acid salt is calcium lactate, lithium lactate, magnesium lactate or lactic acid
The combination of one or more of zinc, wherein calculated with the total volume of thrombin solution, the water-soluble lactic acid salt it is dense
Degree is 1g/L to 20g/L, and the concentration of the water-soluble glutamate is 1g/L to 150g/L.
2. thrombin solution according to claim 1, wherein the water-soluble glutamate be sodium glutamate, potassium glutamate or its
Combination.
3. thrombin solution according to claim 1, wherein the buffer is 4- hydroxyethyl piperazineethanesulfonic acids, citric acid, phosphorus
Acid, acetic acid, imidazoles, 3- (N- morpholines) third sulphur, two (2- ethoxys) imido grpup three (methylol) methane, trihydroxy methyl amino first
One in alkane, 3- (N- morpholinyls) -2- hydroxy-propanesulfonic acids, N- (2- acetylaminos)-imino-diacetic acetic acid or 2-morpholine ethane sulfonic acid
Kind or two or more combinations.
4. thrombin solution according to claim 1, wherein the surfactant is anionic surfactant, cation
The combination of one or more of property surfactant, zwitterionic surfactant or nonionic surfactant.
5. thrombin solution according to claim 4, wherein the anionic surfactant is lauryl sodium sulfate, ten
Dialkyl sulfonates, dodecyl-N- sodium sarcosinates, sodium taurocholate, NaTDC or one kind in sodium taurodeoxycholate or
Two or more combinations.
6. thrombin solution according to claim 4, wherein the cationic surfactant is cetyl trimethyl bromine
Change the combination of one or more of ammonium, four decyl ammonium bromides or dodecanyl pyridinium chloride.
7. thrombin solution according to claim 4, wherein the zwitterionic surfactant is 3- [(3- courages amido propyl)
Dimethyl ammonium] -1- propane sulfonic acid salt, 3- [(3- courages amido propyl) dimethyl ammonium] -2- hydroxyl -1- propane sulfonic acid salt, palmityl haemolysis
The combination of one or more of lecithin or dodecyl-N- glycine betaines or dodecyl-Beta-alanine.
8. thrombin solution according to claim 4, wherein the nonionic surfactant is octyl glucoside, heptyl sulphur
For glucoside, capryl-N-METHYL-ALPHA-L-GLUCOSAMINE, polyoxyethylene lauryl ether, seven methylhexyl ether of polyoxyethylene, polyoxyethylene
Isooctyl phenyl ether, ethylene nonyl phenyl ether, polyoxyethylene fatty acid ester, sucrose fatty ester or polyethylene oxide sorb
The combination of one or more of sugar alcohol ester.
9. thrombin solution according to claim 1, wherein the preservative is Sodium azide, Ciprofloxacin, propionic acid or benzoic acid
The combination of one or more of sodium.
10. a kind of reagent, including according to the thrombin solution of any one of claim 1 to 9.
11. reagent according to claim 10, for the reagent for detecting fibrinogen or thrombin time.
12. a kind of kit, it includes the reagents according to claim 10 or 11.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610874963.3A CN106434610B (en) | 2016-09-30 | 2016-09-30 | Thrombin solution |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610874963.3A CN106434610B (en) | 2016-09-30 | 2016-09-30 | Thrombin solution |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106434610A CN106434610A (en) | 2017-02-22 |
| CN106434610B true CN106434610B (en) | 2018-08-31 |
Family
ID=58172844
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610874963.3A Active CN106434610B (en) | 2016-09-30 | 2016-09-30 | Thrombin solution |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106434610B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPWO2020067171A1 (en) * | 2018-09-25 | 2021-10-28 | 積水メディカル株式会社 | Blood coagulation time measurement method |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0302754B1 (en) * | 1987-08-05 | 1993-11-03 | Green Cross Corporation | Stable aqueous thrombin solution |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE69316293T2 (en) * | 1992-09-09 | 1998-08-27 | Tokuyama Corp | Method for determining fibrinogen, dry reagent therefor and method for producing the same |
| CN1243302C (en) * | 2004-01-19 | 2006-02-22 | 庄德重 | Consonant and vowel font code Chinese characters input method |
-
2016
- 2016-09-30 CN CN201610874963.3A patent/CN106434610B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0302754B1 (en) * | 1987-08-05 | 1993-11-03 | Green Cross Corporation | Stable aqueous thrombin solution |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106434610A (en) | 2017-02-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106350499B (en) | The stabilizer of thrombin solution | |
| CN103197083B (en) | D-dimer quality control product and preparation method thereof | |
| CN106350500B (en) | Thrombin solution | |
| CN103163307A (en) | Blood coagulation quality control product and preparing method thereof | |
| CN104630325B (en) | A kind of liquid-type fibrinogen detection reagent and preparation method thereof | |
| CN109541242A (en) | A kind of kaolin reagent quality-control product and its preparation method and application | |
| JP4399615B2 (en) | Stabilizing means and composition for thrombin | |
| WO2002097391A2 (en) | Automated method of correcting for interference of mean cell hemoglobin concentration | |
| CN106434610B (en) | Thrombin solution | |
| CN106337044B (en) | Thrombin solution | |
| CN101526538A (en) | Method for preparing liquid porcellanite APTT reagent | |
| CN110714051B (en) | Protein C activity determination kit | |
| CN106645665A (en) | Thrombin time detection reagent | |
| US9733262B2 (en) | Method for measuring blood coagulation | |
| CN106480007B (en) | The stabilizer of thrombin solution | |
| CN111638374B (en) | In-vitro diagnostic kit for determining prothrombin time | |
| CN106350501B (en) | The stabilizer of thrombin solution | |
| CN114544983A (en) | Preparation method of normal value quality control product for blood coagulation and platelet function analyzer | |
| CN110777137B (en) | Buffer composition for purifying tissue factor, purification preparation, PT detection composition and PT detection kit | |
| CN112557665A (en) | Confirmation reagent for lupus anticoagulant and preparation method thereof | |
| CN115704749A (en) | Kit for preparing glycosylated hemoglobin quality control product and preparation method thereof | |
| Baumgarten et al. | Changes of platelet function and blood coagulation during short-term storage of CPDA-1-stabilised ovine blood | |
| JP5425062B2 (en) | Method for measuring glycoalbumin and the like using a control sample containing D-mannitol | |
| CN112240931A (en) | Immunity turbidimetry kit | |
| Jensen et al. | Influence of freeze-drying on the clotting properties of fibrinogen in plasma |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information | ||
| CB02 | Change of applicant information |
Address after: High tech Zone Chengdu city Sichuan province 611731 rivers Road No. 16 Applicant after: Mike biological Limited by Share Ltd Address before: High tech Zone Chengdu city Sichuan province 611731 rivers Road No. 16 Applicant before: Sichuan Maker Biological Science and Technology Co., Ltd. |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |