CN112114132A - Carrier fixed with nitrocellulose membrane, preparation method and application thereof - Google Patents
Carrier fixed with nitrocellulose membrane, preparation method and application thereof Download PDFInfo
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- CN112114132A CN112114132A CN202010886730.1A CN202010886730A CN112114132A CN 112114132 A CN112114132 A CN 112114132A CN 202010886730 A CN202010886730 A CN 202010886730A CN 112114132 A CN112114132 A CN 112114132A
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- nitrocellulose membrane
- carrier
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- 239000000020 Nitrocellulose Substances 0.000 title claims abstract description 69
- 229920001220 nitrocellulos Polymers 0.000 title claims abstract description 69
- 239000012528 membrane Substances 0.000 title claims abstract description 67
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- 238000005507 spraying Methods 0.000 claims abstract description 37
- 238000000034 method Methods 0.000 claims abstract description 23
- 239000011259 mixed solution Substances 0.000 claims abstract description 21
- 239000003960 organic solvent Substances 0.000 claims abstract description 16
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 24
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 20
- 239000000203 mixture Substances 0.000 claims description 10
- 238000004519 manufacturing process Methods 0.000 claims description 8
- 238000000889 atomisation Methods 0.000 claims description 2
- 239000002184 metal Substances 0.000 claims description 2
- 238000010186 staining Methods 0.000 claims description 2
- -1 acryl Chemical group 0.000 claims 1
- SWXVUIWOUIDPGS-UHFFFAOYSA-N diacetone alcohol Natural products CC(=O)CC(C)(C)O SWXVUIWOUIDPGS-UHFFFAOYSA-N 0.000 claims 1
- 229920001296 polysiloxane Polymers 0.000 claims 1
- FJWGYAHXMCUOOM-QHOUIDNNSA-N [(2s,3r,4s,5r,6r)-2-[(2r,3r,4s,5r,6s)-4,5-dinitrooxy-2-(nitrooxymethyl)-6-[(2r,3r,4s,5r,6s)-4,5,6-trinitrooxy-2-(nitrooxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-3,5-dinitrooxy-6-(nitrooxymethyl)oxan-4-yl] nitrate Chemical compound O([C@@H]1O[C@@H]([C@H]([C@H](O[N+]([O-])=O)[C@H]1O[N+]([O-])=O)O[C@H]1[C@@H]([C@@H](O[N+]([O-])=O)[C@H](O[N+]([O-])=O)[C@@H](CO[N+]([O-])=O)O1)O[N+]([O-])=O)CO[N+](=O)[O-])[C@@H]1[C@@H](CO[N+]([O-])=O)O[C@@H](O[N+]([O-])=O)[C@H](O[N+]([O-])=O)[C@H]1O[N+]([O-])=O FJWGYAHXMCUOOM-QHOUIDNNSA-N 0.000 abstract description 62
- 230000008569 process Effects 0.000 abstract description 3
- 239000007921 spray Substances 0.000 abstract description 3
- 230000000052 comparative effect Effects 0.000 description 20
- 238000003825 pressing Methods 0.000 description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical group O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- 239000000741 silica gel Substances 0.000 description 4
- 229910002027 silica gel Inorganic materials 0.000 description 4
- 230000009471 action Effects 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 239000011261 inert gas Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 238000004140 cleaning Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- NIXOWILDQLNWCW-UHFFFAOYSA-N acrylic acid group Chemical group C(C=C)(=O)O NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000036046 immunoreaction Effects 0.000 description 1
- HWYHZTIRURJOHG-UHFFFAOYSA-N luminol Chemical compound O=C1NNC(=O)C2=C1C(N)=CC=C2 HWYHZTIRURJOHG-UHFFFAOYSA-N 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000005096 rolling process Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
Images
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Biotechnology (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
Abstract
The embodiment of the invention discloses a carrier fixed with a nitrocellulose membrane, a preparation method and an application thereof, wherein the preparation method comprises the following steps: dissolving a nitrocellulose membrane in an organic solvent to obtain a mixed solution; uniformly spraying the mixed solution on the surface of a carrier to obtain the carrier fixed with the nitrocellulose membrane; according to the invention, the nitrocellulose membrane is dissolved in the organic solvent and then sprayed, so that the damage to the nitrocellulose membrane can be avoided, the operation of fixing the nitrocellulose membrane on the surface of a carrier in the prior art is avoided, the operation steps are saved, and the damage caused in the fixing process of the nitrocellulose membrane is reduced; in addition, the method can spray the nitrocellulose membrane on the carrier in any shape and any thickness to meet various requirements, and the method also improves the bonding strength of the nitrocellulose membrane and the carrier and can prevent the cellulose nitrate membrane from curling.
Description
Technical Field
The embodiment of the invention relates to the technical field of immunoreaction, in particular to a carrier fixed with a nitrocellulose membrane, a preparation method and application thereof.
Background
The nitrocellulose membrane is one of the most important consumable items in biological tests, is one of the most extensive specific transfer mediums, has strong binding capacity to protein, and is suitable for various color development methods, including isotope, chemiluminescence (Luminol), conventional color development, dyeing and fluorescence color development; has low background and high signal-to-noise ratio, and is a basic carrier in an immune response system.
The existing competitive product technology directly selects the required nitrocellulose membrane, combines with a supporting gasket through glue, and fixes the nitrocellulose membrane on the surface of a carrier; however, this method has the following problems: the nitrocellulose membrane needs to be soaked in deionized water to discharge bubbles in the membrane, and then is balanced in an electrophoresis buffer solution for several minutes, so that the operation is complex and the time is long; in addition, nitrocellulose membranes are brittle and easy to roll, require extra care in handling, and are not suitable for applications requiring repeated cleaning.
Disclosure of Invention
Therefore, the embodiment of the invention provides a preparation method of a carrier fixed with a nitrocellulose membrane, which aims to solve the problems of complex operation, easy rolling and unsuitability for repeated cleaning and use caused by the fact that the nitrocellulose membrane needs to be directly fixed on the surface of the carrier in the prior art.
In order to achieve the above object, the embodiments of the present invention provide the following technical solutions:
according to a first aspect of embodiments of the present invention, there is provided a method of producing a nitrocellulose membrane-immobilized carrier, the method comprising the steps of:
(a) dissolving a nitrocellulose membrane in an organic solvent to obtain a mixed solution;
(b) and uniformly spraying the mixed solution on the surface of the carrier to obtain the carrier fixed with the nitrocellulose membrane.
According to the invention, the nitrocellulose membrane is dissolved in the organic solvent and then sprayed, so that the damage to the nitrocellulose membrane (such as reduction of the flatness and scratch generation of the nitrocellulose membrane) can be avoided, the operation of fixing the nitrocellulose membrane on the surface of a carrier in the prior art is avoided, the operation steps are saved, and the damage caused in the fixing process of the nitrocellulose membrane is reduced; in addition, the method can spray the nitrocellulose membrane on the carrier in any shape and any thickness to meet various requirements, and the method also improves the bonding strength of the nitrocellulose membrane and the carrier and can prevent the cellulose nitrate membrane from curling.
Further, the mass concentration of the nitrocellulose membrane in the mixed solution is 0.003-0.007 g/ml; according to the invention, the compactness of the membrane can be controlled by limiting the concentration of the nitrocellulose membrane, so that the mass transfer rate of the membrane is influenced.
Further, the organic solvent is a mixture of acetone and isopropanol; the volume ratio of acetone to isopropanol in the mixture is 2: 5-7; the invention ensures that the film is light, thin and uniform, and has thicker skin layer and lower porosity by selecting the specific organic solvent and limiting the content.
Further, the spraying is as follows: the mixed solution is atomized according to the flow rate of 480-. The invention can control the thickness and the smoothness of the membrane by controlling the flow rate of the mixed solution.
Furthermore, the directional spraying is that transverse spraying is firstly carried out in a spraying area, and then longitudinal spraying is carried out. By controlling the spraying mode, the smoothness of the formed nitrocellulose membrane can be guaranteed.
Further, the directional spraying is controlled through a jig.
Further, the spraying is carried out by adopting a precise electromagnetic valve.
Further, the atomization is performed using an ultrasonic atomizer.
Further, the carrier is a silica gel sheet, a rubber sheet, a metal panel or an acrylic sheet.
According to a second aspect of embodiments of the present invention, there is provided a nitrocellulose membrane-immobilized carrier produced by the above production method.
According to a third aspect of embodiments of the present invention, there is provided a use of the above-described nitrocellulose membrane-immobilized carrier in isotope, chemiluminescence, conventional coloration, staining, or fluorography.
The embodiment of the invention has the following advantages:
according to the invention, the nitrocellulose membrane is dissolved in the organic solvent and then sprayed, so that the damage to the nitrocellulose membrane can be avoided, the operation of fixing the nitrocellulose membrane on the surface of a carrier in the prior art is avoided, the operation steps are saved, and the damage caused in the fixing process of the nitrocellulose membrane is reduced; in addition, the method can spray the nitrocellulose membrane on the carrier in any shape and any thickness to meet various requirements, and the method also improves the bonding strength of the nitrocellulose membrane and the carrier and can prevent the cellulose nitrate membrane from curling.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. It should be apparent that the drawings in the following description are merely exemplary, and that other embodiments can be derived from the drawings provided by those of ordinary skill in the art without inventive effort.
FIG. 1 shows the result of spotting the pressed sample of the product prepared in example 3 provided in Experimental example 2 of the present invention;
FIG. 2 shows the result of spotting the product by pressing the cartridge prepared in comparative example 1 provided in Experimental example 2 of the present invention;
FIG. 3 shows the result of spotting the product by pressing the cartridge prepared in comparative example 2 provided in Experimental example 2 of the present invention;
FIG. 4 shows the result of spotting the product by pressing the cartridge prepared in comparative example 3 provided in Experimental example 2 of the present invention.
Detailed Description
The present invention is described in terms of particular embodiments, other advantages and features of the invention will become apparent to those skilled in the art from the following disclosure, and it is to be understood that the described embodiments are merely exemplary of the invention and that it is not intended to limit the invention to the particular embodiments disclosed. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The following examples used the following raw materials and equipment:
carrier: a silica gel sheet; obtained by ordinary market;
precision electromagnetic valve: model kd1206pts 1; a manufacturer sunon;
precision sample injector: model 82520; the manufacturer: hamilton;
ultrasonic atomizer: model number 06-05108; manufacturer SONO-TEK.
Example 1
This embodiment is a method for preparing a carrier on which a nitrocellulose membrane is immobilized, the method comprising the steps of:
(a) dissolving 0.28g of cellulose nitrate membrane in 40ml of organic solvent to obtain a mixed solution, wherein the organic solvent is a mixture of acetone and isopropanol, and the volume ratio of the acetone to the isopropanol in the mixture is 2: 7;
(b) atomizing the mixed solution by an ultrasonic atomizer at a flow rate of 480 mul/min, and then directionally spraying the atomized mixed solution to a spraying area on the surface of a carrier under the action of a precise electromagnetic valve through the guidance of inert gas to obtain the carrier fixed with the nitrocellulose membrane, wherein the flow rate is controlled by a precise sample injector, and the directional spraying is that transverse spraying is firstly carried out in the spraying area, and then longitudinal spraying is carried out.
Example 2
This embodiment is a method for preparing a carrier on which a nitrocellulose membrane is immobilized, the method comprising the steps of:
(a) dissolving 0.12g of cellulose nitrate membrane in 40ml of organic solvent to obtain a mixed solution, wherein the organic solvent is a mixture of acetone and isopropanol, and the volume ratio of the acetone to the isopropanol in the mixture is 2: 5;
(b) atomizing the mixed solution by an ultrasonic atomizer according to the flow rate of 520 mul/min, and then directionally spraying the atomized mixed solution to a spraying area on the surface of a carrier under the action of a precise electromagnetic valve through the guidance of inert gas to obtain the carrier fixed with the nitrocellulose membrane, wherein the flow rate is controlled by a precise sample injector, and the directional spraying is that transverse spraying is firstly carried out in the spraying area, and then longitudinal spraying is carried out.
Example 3
This embodiment is a method for preparing a carrier on which a nitrocellulose membrane is immobilized, the method comprising the steps of:
(a) dissolving 0.2g of cellulose nitrate membrane in 40ml of organic solvent to obtain a mixed solution, wherein the organic solvent is a mixture of acetone and isopropanol, and the volume ratio of the acetone to the isopropanol in the mixture is 2: 6;
(b) atomizing the mixed solution by an ultrasonic atomizer according to the flow rate of 500 mul/min, and then directionally spraying the atomized mixed solution to a spraying area on the surface of a carrier under the action of a precise electromagnetic valve through the guidance of inert gas to obtain the carrier fixed with the nitrocellulose membrane, wherein the flow rate is controlled by a precise sample injector, and the directional spraying is that transverse spraying is firstly carried out in the spraying area, and then longitudinal spraying is carried out.
Comparative example 1
The present comparative example is a method for producing a nitrocellulose membrane-immobilized carrier, which is substantially the same as the production method in example 3 except that the organic solvent is isopropyl alcohol.
Comparative example 2
This comparative example is a production method of a nitrocellulose membrane-immobilized carrier, which is substantially the same as that in example 3 except that the amount of the nitrocellulose membrane added is 0.4 g.
Comparative example 3
This comparative example is a production method of a nitrocellulose membrane-immobilized carrier, which is substantially the same as that in example 3 except that the flow rate of the mixed solution in step (b) is 550. mu.l/min.
Experimental example 1
In this experimental example, the products and the nitrocellulose membranes prepared in example 3 and comparative examples 1 to 3 were selected, the water contact angle detection was performed on the silica gel sheet membrane spraying area and the nitrocellulose membrane, 3 samples were detected, and the average value was calculated, and the calculation results are shown in table 1:
TABLE 1
| Group of | Nitrocellulose membrane | Example 3 | Comparative example 1 | Comparative example 2 | Comparative example 3 |
| Contact angle (°) | ＞90 | 107.053 | 88.155 | 85.324 | 86.364 |
As can be seen from Table 1: compared with the comparison examples 1-3, the nitrocellulose membrane prepared by the invention has the water contact angle of more than 90 degrees, is hydrophobic, and the comparison examples 1-3 are all less than 90 degrees, which is not qualified.
Experimental example 2
Selecting the products prepared in the example 3 and the comparative examples 1-3 respectively; and pressing and spotting the selected silica gel sheet on the film spraying area and the nitrocellulose membrane, and taking a picture by a camera, wherein the shooting result is shown in figures 1-4:
FIG. 1 shows the result of spotting the pressed cells of the product prepared in example 3; FIG. 2 shows the result of spotting the product prepared in comparative example 1 by pressing; FIG. 3 shows the result of spotting the product prepared in comparative example 2; FIG. 4 shows the result of spotting the product prepared in comparative example 3 by pressing;
as can be seen from fig. 1-4: the product prepared in comparative example 1 was slightly soluble in the nitrocellulose membrane due to the choice of solvent and could not be detected; in comparative example 2, when the amount of the nitrocellulose membrane added was too large, the background value of the result was too bright to be detected; the product prepared in comparative example 3 interfered the detection result when the flow rate was too fast; the product prepared by the invention has excellent detection effect.
Although the invention has been described in detail above with reference to a general description and specific examples, it will be apparent to one skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (10)
1. A preparation method of a carrier fixed with a nitrocellulose membrane is characterized by comprising the following steps:
(a) dissolving a nitrocellulose membrane in an organic solvent to obtain a mixed solution;
(b) and uniformly spraying the mixed solution on the surface of the carrier to obtain the carrier fixed with the nitrocellulose membrane.
2. The production method according to claim 1, wherein the mass concentration of the nitrocellulose membrane in the mixed solution is 0.003 to 0.007 g/ml.
3. The production method according to claim 1, wherein the organic solvent is a mixture of acetone and isopropyl alcohol; the volume ratio of the acetone to the isopropanol in the mixture is 2: 5-7.
4. The method of claim 1, wherein the spraying is: the mixed solution is atomized according to the flow rate of 480-.
5. The preparation method according to claim 4, wherein the directional spraying is performed by firstly performing transverse spraying in a spraying area and then performing longitudinal spraying.
6. The method of claim 4, wherein the spraying is performed using a precision solenoid valve.
7. The method of claim 4, wherein the atomization is performed using an ultrasonic atomizer.
8. The method according to claim 1, wherein the carrier is a silicone sheet, a rubber sheet, a metal panel, or an acryl sheet.
9. A carrier having a nitrocellulose membrane immobilized thereon, which is produced by the production method according to any one of claims 1 to 8.
10. Use of the nitrocellulose membrane-immobilized carrier of claim 9 in isotope, chemiluminescence, conventional coloration, staining or fluorogenic coloration.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010886730.1A CN112114132A (en) | 2020-08-28 | 2020-08-28 | Carrier fixed with nitrocellulose membrane, preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010886730.1A CN112114132A (en) | 2020-08-28 | 2020-08-28 | Carrier fixed with nitrocellulose membrane, preparation method and application thereof |
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| Publication Number | Publication Date |
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| CN112114132A true CN112114132A (en) | 2020-12-22 |
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| CN202010886730.1A Pending CN112114132A (en) | 2020-08-28 | 2020-08-28 | Carrier fixed with nitrocellulose membrane, preparation method and application thereof |
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