CN113607962B - Preservation solution for cTnI antibody coated magnetic beads and preparation method thereof - Google Patents

Preservation solution for cTnI antibody coated magnetic beads and preparation method thereof Download PDF

Info

Publication number
CN113607962B
CN113607962B CN202110905348.5A CN202110905348A CN113607962B CN 113607962 B CN113607962 B CN 113607962B CN 202110905348 A CN202110905348 A CN 202110905348A CN 113607962 B CN113607962 B CN 113607962B
Authority
CN
China
Prior art keywords
concentration
preservation solution
magnetic beads
coated magnetic
ctni antibody
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
Application number
CN202110905348.5A
Other languages
Chinese (zh)
Other versions
CN113607962A (en
Inventor
周倩
毛善检
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Sinocare Inc
Original Assignee
Sinocare Inc
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Sinocare Inc filed Critical Sinocare Inc
Priority to CN202110905348.5A priority Critical patent/CN113607962B/en
Priority to PCT/CN2021/117122 priority patent/WO2023010647A1/en
Publication of CN113607962A publication Critical patent/CN113607962A/en
Application granted granted Critical
Publication of CN113607962B publication Critical patent/CN113607962B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6887Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids from muscle, cartilage or connective tissue
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54313Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
    • G01N33/54326Magnetic particles
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/46Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
    • G01N2333/47Assays involving proteins of known structure or function as defined in the subgroups
    • G01N2333/4701Details
    • G01N2333/4712Muscle proteins, e.g. myosin, actin, protein
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00Detection or diagnosis of diseases
    • G01N2800/32Cardiovascular disorders
    • G01N2800/324Coronary artery diseases, e.g. angina pectoris, myocardial infarction
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02ATECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The application discloses preservation solution of cTnI antibody coating magnetic bead, solvent is water, its characterized in that includes: a buffer salt; a protein; a surfactant; an inorganic salt; the protective agent is cetostearyl alcohol polyether-25; a preservative; the pH of the preservation solution is 7.0-8.0. The application also provides a preparation method of the preservation solution of the cTnI antibody coated magnetic beads. The preservation solution of the cTnI antibody coated magnetic beads can effectively improve the stability of the cTnI antibody coated magnetic beads in the storage and use processes; in particular, the addition of the protective agent can significantly improve the stability, sensitivity and linear gradient of the reagent. And the preservation solution of the cTnI antibody coated magnetic beads has the advantages of simple components, simple and convenient preparation, low cost and convenient realization of mass production.

Description

Preservation solution for cTnI antibody coated magnetic beads and preparation method thereof
Technical Field
The application relates to the technical field of detection reagents, in particular to preservation solution of cTnI antibody coated magnetic beads and a preparation method thereof.
Background
Troponin (Tn) complex is composed of troponin I, troponin T and troponin C and is able to regulate muscle contraction. Cardiac troponin I (cardiactroponin I, cTnI) is an isoform of troponin I found in cardiomyocytes and has a high degree of myocardial specificity and can be used to identify clinical situations associated with myocardial and skeletal muscle injury.
Acute myocardial infarction (Acute Myocardial Infarction, AMI) is one of the major diseases that are currently life threatening in humans. When myocardial cells are damaged by myocardial infarction or ischemia, cTnI rapidly penetrates through myocardial cell membranes into blood. Therefore, in the early stage of onset, the cTnI and the change trend thereof in human blood can be rapidly, sensitively and accurately measured, and the method has important clinical significance for diagnosis and treatment of acute myocardial infarction.
The cTnI is used as a first-choice marker of myocardial injury, a common method for clinical detection at present is a chemiluminescent immunoassay method, and a magnetic particle chemiluminescent method is mainly adopted at present. The magnetic bead coated antibody plays an important role in the chemiluminescent detection reagent, so that the stability of the magnetic bead coated antibody lays a solid foundation for the high-quality chemiluminescent reagent.
At present, the stability of the magnetic bead coating antibody is improved, mainly starting from the optimization of the magnetic bead coating process or the optimization of the formula. However, the former is relatively complex in process, while the latter usually adopts commercial magnetic bead storage liquid, is relatively expensive, and has certain batch-to-batch difference to influence the reagent performance.
Therefore, how to provide a preservation solution for improving the use and storage stability of cTnI antibody coated magnetic beads, improving the storage stability, reducing the batch-to-batch difference and simultaneously reducing the cost is a technical problem to be solved by the person skilled in the art.
Disclosure of Invention
In order to solve the technical problems, a first object of the present invention is to provide a preservation solution of cTnI antibody coated magnetic beads; the second object of the invention is to provide a method for preparing preservation solution of the cTnI antibody-coated magnetic beads; the preservation solution of the cTnI antibody coated magnetic beads can effectively improve the stability of the cTnI antibody coated magnetic beads in the storage and use processes, and protective agents such as sugar, alcohol, lipid and the like are not required to be added; in particular, the addition of the protective agent can significantly improve the stability, sensitivity and linear gradient of the reagent. And the preservation solution of the cTnI antibody coated magnetic beads has the advantages of simple components, simple and convenient preparation, low cost and convenient realization of mass production.
The technical scheme provided by the invention is as follows:
a preservation solution of cTnI antibody coated magnetic beads, the solvent being water, comprising:
a buffer salt;
a protein;
a surfactant;
an inorganic salt;
the protective agent is cetostearyl alcohol polyether-25;
a preservative;
the pH of the preservation solution is 7.0-8.0.
Preferably, it comprises:
20-100mmol/L of buffer salt;
2-20g/L of protein;
0.5-5g/L of surfactant;
0.1 to 0.5mol/L of inorganic salt;
the protective agent is cetostearyl alcohol polyether-25-10 g/L;
0.5-5g/L preservative;
the pH of the preservation solution is 7.0-8.0.
Preferably, the buffer salt is specifically any one of Tris, MES, MOPS, HEPES; and/or the number of the groups of groups,
the concentration of the buffer salt is 50-60mmol/L.
Preferably, the protein is specifically any one or more of bovine serum albumin, casein, gelatin protein.
Preferably, the protein is bovine serum albumin, and the concentration of the bovine serum albumin is 10-15g/L.
Preferably, the surfactant is any one or more of triton x-100, triton x-405, tween 20 and tween 80; and/or the number of the groups of groups,
the concentration of the surfactant is 1-2g/L.
Preferably, the inorganic salt is specifically a combination of sodium chloride, magnesium chloride, zinc chloride; or the ionic salt is a combination of potassium chloride, magnesium chloride and zinc chloride; the concentration of sodium chloride is 0.1-0.3 mol/L; the concentration of magnesium chloride is 0.5-5mmol/L; the concentration of zinc chloride is 0.05-1mmol/L; the concentration of the potassium chloride is 0.1-0.3 mol/L.
Preferably, in the inorganic salt combination, the concentration of sodium chloride is 0.1-0.2 mol/L; the concentration of magnesium chloride is 2.5-5 mmol/L; the concentration of zinc chloride is 0.1-0.5 mmol/L; the concentration of the potassium chloride is 0.1-0.2 mol/L.
More preferably, the inorganic salt composition used is: sodium chloride 0.15mol/L, magnesium chloride 5mmol/L, zinc chloride 0.1mmol/L;
or 0.15mol/L of potassium chloride, 5mmol/L of magnesium chloride and 0.1mmol/L of zinc chloride;
or, 0.15mol/L of sodium chloride, 2.5mmol/L of magnesium chloride and 00.5mmol/L of zinc chloride;
or 0.15mol/L of potassium chloride, 2.5mmol/L of magnesium chloride and 00.5mmol/L of zinc chloride.
Preferably, the preservative is specifically any one or more of sodium azide, PC300, chloramphenicol and gentamicin; and/or the number of the groups of groups,
the concentration of the preservative is 1-2g/L.
Preferably, the preservation solution is pH-adjusted using an inorganic acid or an inorganic base. Among them, the inorganic acid is preferably hydrochloric acid, and the inorganic base is preferably sodium hydroxide.
A preparation method of preservation solution of cTnI antibody coated magnetic beads comprises the following steps:
respectively adding buffer salt, protein, surfactant, inorganic salt, cetostearyl alcohol polyether-25 serving as a protective agent and preservative into water, and uniformly mixing so that the concentration of the buffer salt is 20-100mmol/L and the concentration of the protein is 2-20g/L; the concentration of the surfactant is 0.5-5g/L; the concentration of the inorganic salt is 0.1-0.5mol/L; the concentration of the protecting agent cetostearyl alcohol polyether-25 is 1-10g/L, and the concentration of the preservative is 0.5-5g/L; and adjusting the pH to 7.0-8.0;
constant volume and filtering. According to the preparation method of the preservation solution of the cTnI antibody coated magnetic beads, which is provided by the application, the sequence of adding buffer salt, protein, surfactant, inorganic salt, preservative cetostearyl alcohol polyether-25 is not required, each substance is only required to be added independently and uniformly mixed, then the next is added, and finally, the pH value, the constant volume and the filtration are adjusted.
The preservation solution of the cTnI antibody coated magnetic beads can effectively improve the stability of the cTnI antibody coated magnetic beads in the storage and use processes, and protective agents such as sugar, alcohol, lipid and the like are not required to be added; in particular, the addition of the protective agent can significantly improve the stability, sensitivity and linear gradient of the reagent. And the preservation solution of the cTnI antibody coated magnetic beads has the advantages of simple components, simple and convenient preparation, low cost and convenient realization of mass production.
Drawings
In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings that are required to be used in the embodiments or the description of the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments described in the present application, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a graph showing the results of a dose response curve test in an embodiment of the present invention;
Detailed Description
In order to better understand the technical solutions in the present application, the following description will clearly and completely describe the technical solutions in the embodiments of the present application with reference to the drawings in the embodiments of the present application, and it is obvious that the described embodiments are only some embodiments of the present application, not all embodiments. All other embodiments, which can be made by one of ordinary skill in the art based on the embodiments herein without making any inventive effort, shall fall within the scope of the present application.
It will be understood that when an element is referred to as being "fixed" or "disposed" on another element, it can be directly on the other element or be indirectly on the other element; when an element is referred to as being "connected to" another element, it can be directly connected to the other element or be indirectly connected to the other element.
It is to be understood that the terms "length," "width," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," and the like indicate or are based on the orientation or positional relationship shown in the drawings, merely to facilitate description of the present application and simplify description, and do not indicate or imply that the devices or elements referred to must have a particular orientation, be configured and operated in a particular orientation, and therefore should not be construed as limiting the present application.
Furthermore, the terms "first," "second," and the like, are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include one or more such feature. In the description of the present application, the meaning of "a plurality" or "a number" is two or more, unless explicitly defined otherwise.
It should be understood that the structures, proportions, sizes, etc. shown in the drawings are for illustration purposes only and should not be construed as limiting the scope of the present disclosure, since any structural modifications, proportional changes, or dimensional adjustments made by those skilled in the art should not be made in the present disclosure without affecting the efficacy or achievement of the present disclosure.
The embodiment of the application is written in a progressive manner.
Example 1
A preservation solution of cTnI antibody coated magnetic beads, the solvent being pure water, comprising:
buffer salt: tris 50mmol/L;
protein: bovine serum albumin 10g/L;
and (2) a surfactant: triton x-100 g/L;
inorganic salts including sodium chloride: 0.15mol/L; magnesium chloride: 5.00mmol/L; zinc chloride: 0.10mmol/L;
and (3) a protective agent: cetostearyl alcohol polyether-25 g/L;
preservative: 1g/L of sodium azide and 300 g/L of PC;
the pH of the preservation solution was 7.4.
The preparation method of the preservation solution of the cTnI antibody coated magnetic beads comprises the following steps:
adding buffer salt into water, and uniformly mixing;
adding inorganic salt into water, and uniformly mixing;
adding protein into water, and uniformly mixing;
adding a surfactant into water, and uniformly mixing;
adding a protective agent into water, and uniformly mixing;
adding preservative into water and uniformly mixing;
adjusting the pH to 7.40+/-0.05;
constant volume, and filtering with 0.22um filter element.
The protectant added in the above examples is cetostearyl alcohol polyether-25, cremophor A25; the protective agent added in the subsequent examples was cetostearyl alcohol polyether-25 (Cremophor A25).
Example 2
A preservation solution of cTnI antibody coated magnetic beads, the solvent being pure water, comprising:
buffer salt: tris 50mmol/L;
protein: bovine serum albumin 15g/L;
and (2) a surfactant: triton x-100 g/L;
inorganic salts including sodium chloride: 0.15mol/L; magnesium chloride: 2.50mmol/L; zinc chloride: 0.50mmol/L;
and (3) a protective agent: cetostearyl alcohol polyether-25.10 g/L;
preservative: 1g/L of sodium azide and 300 g/L of PC;
the pH of the preservation solution was 7.4.
The preparation method of the preservation solution of the cTnI antibody coated magnetic beads comprises the following steps:
adding buffer salt into water, and uniformly mixing;
adding protein into water, and uniformly mixing; adding inorganic salt into water, and uniformly mixing;
adding a surfactant into water, and uniformly mixing;
adding a protective agent into water, and uniformly mixing;
adding preservative into water and uniformly mixing;
adjusting the pH to 7.40+/-0.05;
constant volume, and filtering with 0.22um filter element.
Example 3
A preservation solution of cTnI antibody coated magnetic beads, the solvent being pure water, comprising:
buffer salt: tris 50mmol/L;
protein: bovine serum albumin 10g/L;
and (2) a surfactant: triton x-100 g/L;
inorganic salts including sodium chloride: 0.15mmol/L; magnesium chloride: 2.50mmol/L; zinc chloride: 0.50mmol/L;
and (3) a protective agent: cetostearyl alcohol polyether-25 g/L;
preservative: 1g/L of sodium azide and 300 g/L of PC;
the pH of the preservation solution was 7.4.
The preparation method of the preservation solution of the cTnI antibody coated magnetic beads comprises the following steps:
adding buffer salt into water, and uniformly mixing;
adding a surfactant into water, and uniformly mixing;
adding a protective agent into water, and uniformly mixing;
adding inorganic salt into water, and uniformly mixing;
adding protein into water, and uniformly mixing;
adding preservative into water and uniformly mixing;
adjusting the pH to 7.40+/-0.05;
constant volume, and filtering with 0.22um filter element.
Comparative example 1
A preservation solution of cTnI antibody coated magnetic beads, the solvent being pure water, comprising:
buffer salt: tris 50mmol/L;
protein: bovine serum albumin 10g/L;
and (2) a surfactant: triton x-100 g/L;
inorganic salts including sodium chloride: 0.15mol/L; magnesium chloride: 2.50mmol/L; zinc chloride: 0.50mmol/L;
preservative: 1g/L of sodium azide and 300 g/L of PC;
the pH of the preservation solution was 7.4.
The preparation method of the preservation solution of the cTnI antibody coated magnetic beads comprises the following steps:
adding buffer salt into water, and uniformly mixing;
adding inorganic salt into water, and uniformly mixing;
adding protein into water, and uniformly mixing;
adding a surfactant into water, and uniformly mixing;
adding preservative into water and uniformly mixing;
adjusting the pH to 7.40+/-0.05;
constant volume, and filtering with 0.22um filter element.
Comparative example 2
A preservation solution of cTnI antibody coated magnetic beads, the solvent being pure water, comprising:
buffer salt: tris 50mmol/L;
protein: bovine serum albumin 10g/L;
and (2) a surfactant: triton x-100 g/L;
inorganic salts including sodium chloride: 0.15mol/L; magnesium chloride: 2.50mmol/L; zinc chloride: 0.50mmol/L;
preservative: 1g/L of sodium azide and 300 g/L of PC;
and (3) a protective agent: glycerin 50g/L, sucrose 10g/L, polyethylene glycol 4000 1g/L;
the pH of the preservation solution was 7.4.
The preparation method of the preservation solution of the cTnI antibody coated magnetic beads comprises the following steps:
adding buffer salt into water, and uniformly mixing;
adding inorganic salt into water, and uniformly mixing;
adding protein into water, and uniformly mixing;
adding a surfactant into water, and uniformly mixing;
adding a protective agent into water, and uniformly mixing;
adding preservative into water and uniformly mixing;
adjusting the pH to 7.40+/-0.05;
constant volume, and filtering with 0.22um filter element.
Comparative example 3
A preservation solution of cTnI antibody coated magnetic beads, the solvent being pure water, comprising:
buffer salt: tris 50mmol/L;
protein: bovine serum albumin 10g/L;
and (2) a surfactant: triton x-100 g/L;
inorganic salts including sodium chloride: 0.15mol/L; magnesium chloride: 2.50mmol/L; zinc chloride: 0.50mmol/L;
and (3) a protective agent: cetostearyl alcohol polyether-25.5 g/L;
preservative: 1g/L of sodium azide and 300 g/L of PC;
the pH of the preservation solution was 7.4.
The preparation method of the preservation solution of the cTnI antibody coated magnetic beads comprises the following steps:
adding buffer salt into water, and uniformly mixing;
adding inorganic salt into water, and uniformly mixing;
adding protein into water, and uniformly mixing;
adding a surfactant into water, and uniformly mixing;
adding a protective agent into water, and uniformly mixing;
adding preservative into water and uniformly mixing;
adjusting the pH to 7.40+/-0.05;
constant volume, filtering with 0.22um filter element
Comparative example 4
A preservation solution of cTnI antibody coated magnetic beads, the solvent being pure water, comprising:
buffer salt: tris 50mmol/L;
protein: bovine serum albumin 10g/L;
and (2) a surfactant: triton x-100 g/L;
inorganic salts including sodium chloride: 0.15mol/L; magnesium chloride: 2.50mol/L; zinc chloride: 0.50mol/L;
and (3) a protective agent: cetostearyl alcohol polyether-25 g/L;
preservative: 1g/L of sodium azide and 300 g/L of PC;
the pH of the preservation solution was 7.4.
The preparation method of the preservation solution of the cTnI antibody coated magnetic beads comprises the following steps:
adding buffer salt into water, and uniformly mixing;
adding inorganic salt into water, and uniformly mixing;
adding protein into water, and uniformly mixing;
adding a surfactant into water, and uniformly mixing;
adding a protective agent into water, and uniformly mixing;
adding preservative into water and uniformly mixing;
adjusting the pH to 7.40+/-0.05;
constant volume, filtering with 0.22um filter element
The preservation solutions of the cTnI antibody-coated magnetic beads of examples 1 to 3 and comparative examples 1 to 4 were tested.
Experiment one: stability verification
The magnetic bead reagent is prepared by adding cTnI antibody coated magnetic bead mother liquor into the preservation solution.
The reagent is divided into 2 parts, one part is placed at 37 ℃ and aged for 7 days, and the other part is placed at 2-8 ℃ and stored. After 7 days of standing, the same samples were tested simultaneously with two reagents using the reagent at 2-8deg.C as a control. The test results were as follows:
TABLE 1 stability test results
Figure BDA0003200275640000101
From the results of experimental examples 1-3, the working solution obtained by diluting the cTnI antibody coated magnetic beads with the preservation solution is accelerated for 7 days at 37 ℃, and the luminous value is reduced to be within 3.0%; the concentration of the cetostearyl alcohol polyether-25 is in the range of 1-10g/L, and has no obvious influence on stability. As is clear from comparative example 1, the luminous value without the addition of the protective agent was reduced by 20.0% or more. As is clear from comparative example 2, the preservation solution was added with a protective agent such as an alcohol, a saccharide, or a polymer, and the luminescence value was reduced by about 15.0%. As can be seen from comparative examples 3-4, too much or too little cetostearyl alcohol polyether-25 concentration affects reagent stability.
Experiment II: sensitivity verification
The magnetic bead reagent is prepared by adding cTnI antibody coated magnetic bead mother liquor into the preservation solution, and the cTnI reagent is formed by matching with an enzyme marker, a 0 concentration sample is tested for 20 times, a blank limit is calculated, and the test result is as follows:
TABLE 2 sensitivity test results
Figure BDA0003200275640000102
Figure BDA0003200275640000111
From the results of experimental examples 1 to 3, the working solution obtained by diluting cTnI antibody-coated magnetic beads with the preservation solution of the present invention had a blank of 0.002. As is clear from comparative examples 1 to 2, no addition of cetostearyl alcohol polyether-25 protecting agent or other protecting agents such as alcohols, saccharides and polymers has a large influence on the blank space. As can be seen from comparative examples 3-4, too much or too little cetostearyl alcohol polyether-25 concentration is detrimental to blank testing.
The concentrations of examples 1-3 and comparative examples 1-4 were plotted against RLU to give dose response curves as shown in figure 1.
Experiment III: linear gradient validation
The magnetic bead reagent is prepared by adding cTnI antibody coated magnetic bead mother liquor into the preservation solution, and the cTnI reagent is formed by matching with an enzyme marker, and linear gradient test is carried out, and the test result is as follows:
TABLE 3 Linear gradient test results
Figure BDA0003200275640000121
From the results of experimental examples 1-3, the working solution obtained by diluting the cTnI antibody coated magnetic beads with the preservation solution provided by the invention has obvious linear gradient and obvious rising trend within the range of 0-50 ng/mL. As is clear from comparative examples 1-2, the linear gradient was significantly reduced without addition of the cetostearyl alcohol polyether-25 protectant or with addition of other protectants such as alcohols, saccharides, polymers, etc. As can be seen from comparative examples 3-4, too much or too little of cetostearyl alcohol polyether-25 concentration is detrimental to the linear gradient test.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims (8)

1. The preservation solution of the cTnI antibody coated magnetic beads is characterized by comprising the following components:
20-100mmol/L of buffer salt;
2-20g/L of protein;
0.5-5g/L of surfactant;
0.1 to 0.5mol/L of inorganic salt;
the protective agent is cetostearyl alcohol polyether-25-10 g/L;
0.5-5g/L preservative;
the pH value of the preservation solution is 7.0-8.0;
the buffer salt is specifically any one of Tris, MES, MOPS, HEPES;
the protein is specifically any one or more of bovine serum albumin, casein and gelatin protein;
the surfactant is one or more of triton x-100, triton x-405, tween 20 and Tween 80;
the inorganic salt is a combination of sodium chloride, magnesium chloride and zinc chloride; or, a combination of potassium chloride, magnesium chloride and zinc chloride;
the preservation solution does not need sugar, alcohol or lipid protectant.
2. The preservation solution of cTnI antibody-coated magnetic beads according to claim 1,
the concentration of the buffer salt is 50-60mmol/L.
3. The preservative solution for cTnI antibody-coated magnetic beads according to claim 1, wherein the protein is bovine serum albumin, and the concentration of bovine serum albumin is 10-15g/L.
4. The preservation solution of cTnI antibody-coated magnetic beads according to claim 1,
the concentration of the surfactant is 1-2g/L.
5. The preservation solution of cTnI antibody coated magnetic beads according to claim 1, wherein the concentration of sodium chloride in the inorganic salt composition is 0.1 to 0.3mol/L; the concentration of magnesium chloride is 0.5-5mmol/L; the concentration of zinc chloride is 0.05-1mmol/L; the concentration of potassium chloride is 0.1-0.3 mol/L.
6. The preservation solution of cTnI antibody-coated magnetic beads according to claim 5, wherein the concentration of sodium chloride in the inorganic salt composition is 0.1 to 0.2mol/L; the concentration of magnesium chloride is 2.5-5 mmol/L; zinc chloride is 0.1-0.5 mmol/L; the concentration of potassium chloride is 0.1-0.2 mol/L.
7. The preservation solution of cTnI antibody-coated magnetic beads according to claim 1, wherein the preservative is specifically any one or more of sodium azide, PC300, chloramphenicol, and gentamicin; and/or the number of the groups of groups,
the concentration of the preservative is 1-2g/L.
8. A method for preparing a preservation solution of cTnI antibody-coated magnetic beads according to claim 1, comprising the steps of:
respectively adding buffer salt, protein, surfactant, inorganic salt, cetostearyl alcohol polyether-25 serving as a protective agent and preservative into water, and uniformly mixing to ensure that the concentration of the buffer salt is 20-100mmol/L and the concentration of the protein is 2-20g/L; the concentration of the surfactant is 0.5-5g/L; the concentration of the inorganic salt is 0.1-0.5mol/L; the concentration of the protecting agent cetostearyl alcohol polyether-25 is 1-10g/L, and the concentration of the preservative is 0.5-5g/L; and adjusting the pH to 7.0-8.0;
constant volume and filtering.
CN202110905348.5A 2021-08-06 2021-08-06 Preservation solution for cTnI antibody coated magnetic beads and preparation method thereof Active CN113607962B (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
CN202110905348.5A CN113607962B (en) 2021-08-06 2021-08-06 Preservation solution for cTnI antibody coated magnetic beads and preparation method thereof
PCT/CN2021/117122 WO2023010647A1 (en) 2021-08-06 2021-09-08 Ctni antibody-coated magnetic bead preservation solution and preparation method therefor

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110905348.5A CN113607962B (en) 2021-08-06 2021-08-06 Preservation solution for cTnI antibody coated magnetic beads and preparation method thereof

Publications (2)

Publication Number Publication Date
CN113607962A CN113607962A (en) 2021-11-05
CN113607962B true CN113607962B (en) 2023-05-09

Family

ID=78339845

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202110905348.5A Active CN113607962B (en) 2021-08-06 2021-08-06 Preservation solution for cTnI antibody coated magnetic beads and preparation method thereof

Country Status (2)

Country Link
CN (1) CN113607962B (en)
WO (1) WO2023010647A1 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115684581A (en) * 2021-07-28 2023-02-03 北京京东方技术开发有限公司 Preservation solution for magnetic beads and preservation method thereof

Family Cites Families (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0535485B1 (en) * 1991-10-03 1997-07-16 Bayer Corporation Device and method of separating and assaying whole blood
SK53294A3 (en) * 1993-05-07 1995-04-12 Albright & Wilson Concentrated aqueous mixture containing surface active matter and its use
US20040022862A1 (en) * 2000-12-22 2004-02-05 Kipp James E. Method for preparing small particles
GB0623838D0 (en) * 2006-11-29 2007-01-10 Malvern Cosmeceutics Ltd Novel compositions
CN102426246A (en) * 2011-08-31 2012-04-25 内蒙古科慧生物科技有限责任公司 Human troponin I quantitative determination kit and detection method
CN115753300A (en) * 2015-04-20 2023-03-07 文塔纳医疗系统公司 Ink jet deposition of reagents for histological samples
CN108375571B (en) * 2018-02-05 2019-11-26 苏州长光华医生物医学工程有限公司 A kind of magnetic particle reagents and kit for chemiluminescence immunoassay detection
CN108333370A (en) * 2018-02-09 2018-07-27 广州源起健康科技有限公司 A kind of magnetic bead time-resolved fluoroimmunoassay quantitatively detects cTnI kits
CN109061189A (en) * 2018-08-17 2018-12-21 深圳天辰医疗科技有限公司 A kind of Troponin I detection kit and preparation method thereof
CN109646374A (en) * 2019-01-21 2019-04-19 秋臣化妆品有限公司 A kind of herbal mildy wash and preparation method of moisturizing whitening
CN109709323A (en) * 2019-02-15 2019-05-03 深圳上泰生物工程有限公司 High quick cardiac muscle troponin I magnetic microparticle chemiluminescence immunity detection reagent, Preparation method and use
CN112526140A (en) * 2020-11-16 2021-03-19 北京利德曼生化股份有限公司 Magnetic particle chemiluminescence detection kit for determining content of hypersensitive troponin T
CN112698023B (en) * 2020-12-01 2023-12-15 三诺生物传感股份有限公司 Preservation solution of alkaline phosphatase-labeled conjugate and preparation method thereof

Also Published As

Publication number Publication date
WO2023010647A1 (en) 2023-02-09
CN113607962A (en) 2021-11-05

Similar Documents

Publication Publication Date Title
AU570522B2 (en) Stabilization of indicators for detecting enzyme activity
CN113607962B (en) Preservation solution for cTnI antibody coated magnetic beads and preparation method thereof
CN106771133A (en) A kind of composition and its application as enzyme mark compound preservation liquid
EP2324350B1 (en) Reference control for cell by cell analysis
CN112745837B (en) Carbon quantum dot and application thereof
Wrigley et al. Human erythrocyte glucose 6-phosphate dehydrogenase: electron microscope studies on structure and interconversion of tetramers, dimers and monomers
CN112730839B (en) Kit for measuring content of cytokeratin 19 fragments by magnetic particle chemiluminescence method
CN104215772A (en) Heart-type fatty acid binding protein detection kit and making method thereof
DE3311889A1 (en) METHOD FOR IRREVERSIBLE BINDING OF PROTEIN TO POLYSTYROL SURFACES WITH THE PRESERVATION OF BIOLOGICAL ACTIVITY, POLYSTYROL SURFACES OBTAINED AND THEIR USE
CN108709993B (en) Latex-enhanced immunoturbidimetry detection kit and preparation and detection methods thereof
US4486531A (en) Method and reagent for the determination of β-lipoprotein
WO2018216628A1 (en) Protein stabilizer and protein stabilization reagent
CN112698023B (en) Preservation solution of alkaline phosphatase-labeled conjugate and preparation method thereof
Penniston Fluorescence polarization measurement of binding of fluorescein to albumin
Grosicki et al. Isotonic Solutions: III. Amino Acids and Sugars
US20150185236A1 (en) Blood coagulation time prolonging agent
CN107043762A (en) A kind of preserving stabilizer and its store method for improving horseradish peroxidase storage stability
CN116413253A (en) Human papilloma virus detection reagent and application thereof
CN113702647B (en) Human growth differentiation factor 15 instant detection kit, preparation method and application thereof
CN113238060B (en) Kit for predicting or diagnosing myocarditis
Breyer et al. The adsorptive capacity of serum proteins in renal insufficiency.
CN115078733A (en) Method for detecting neutral antibody of cytokine fusion protein
CN112904009A (en) Magnetic microsphere detection kit for detecting glycosylated CD59 and application thereof
CN112240931A (en) Immunity turbidimetry kit
CN111830253A (en) Composition for keeping stability of lipoprotein-associated phospholipase A2 kit and preparation method thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant