CN112816688B - ELISA plate and preparation method and application thereof - Google Patents
ELISA plate and preparation method and application thereof Download PDFInfo
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- CN112816688B CN112816688B CN202110003993.8A CN202110003993A CN112816688B CN 112816688 B CN112816688 B CN 112816688B CN 202110003993 A CN202110003993 A CN 202110003993A CN 112816688 B CN112816688 B CN 112816688B
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- G—PHYSICS
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- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
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- 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
- G01N33/544—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being organic
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Abstract
The invention relates to an ELISA plate and a preparation method and application thereof. The ELISA plate comprises a solid phase carrier and a functional layer modified on the surface of the solid phase carrier, wherein the functional layer comprises a graft copolymer, and a functional monomer of the graft copolymer comprises N-octadecyl acrylamide and/or N- (3-dimethylaminopropyl) methacrylamide. According to the invention, the functional layer is modified on the solid phase carrier of the ELISA plate, so that the surface of the ELISA plate can generate a strong adsorption effect with protein molecules such as antigen or antibody, the effect of coating the protein molecules such as antigen or antibody by the ELISA plate is improved, and the sensitivity of ELISA test is improved.
Description
Technical Field
The invention belongs to the technical field of life science, and relates to an ELISA plate and a preparation method and application thereof.
Background
Enzyme Linked Immunosorbent Assay (ELISA) refers to a qualitative and quantitative detection method in which soluble antigen or antibody is bound to a solid phase carrier such as Polystyrene (PS) and immunoreaction is carried out by utilizing the specific binding of antigen and antibody. ELISA methods are an important component of immunological studies and can be divided into three categories: sandwich method, indirect method and competitive method, wherein the three methods are all required to carry out coating operation, namely coating antigen or antibody on the surface of solid phase carrier.
The binding force between protein biomolecules such as antigens or antibodies and an enzyme label plate solid phase carrier is enhanced, so that the sensitivity and the stability of an ELISA test can be effectively improved. Therefore, in recent years, the selection of solid phase carriers for microplate and the surface modification of the solid phase carriers have become a focus of attention for researchers in this field. Common methods for modifying the surface of the solid phase carrier of the ELISA plate comprise coating a functional layer, chemical modification, ultraviolet irradiation or gamma ray irradiation and the like.
CN103275272A discloses a preparation method of a 96-well enzyme label plate AOZ molecularly imprinted polymeric membrane, which uses template molecule AOZ, functional monomer 2-vinylpyridine and cross-linking agent ethylene glycol dimethacrylate to form a 'memory cavity' with the size, shape and functional groups matched with the template molecules on the surface of the 96-well enzyme label plate, wherein the 'memory cavity' can generate specific affinity action with the template molecules to be separated in a mixture, but the method only improves the specificity, does not improve the adsorption action of the enzyme label plate on protein molecules, can only aim at a specific target substance, and has a small application range.
CN105418955A discloses a method for modifying the surface of polyvinyl chloride, which uses a polyvinyl chloride surface modifier containing a radical initiator and a polyvinyl chloride surface modifying molecule to modify the surface of polyvinyl chloride, so as to enhance the hydrophilic property of the polyvinyl chloride surface, but not enhance the adsorption effect on protein.
In summary, the modified material obtained by the existing modification method still has the problems of low functional layer strength, poor stability, single function, insufficient controllability and consistency of the processing process and the like, which may affect the accuracy, sensitivity and stability of the ELISA test and hinder the application and development of ELISA in different fields. Therefore, research and development are urgently needed for a surface modification method for improving the strength and stability of the solid phase carrier functional layer and for adsorption of proteins.
Disclosure of Invention
Aiming at the defects and actual requirements of the prior art, the invention provides an ELISA plate and a preparation method and application thereof, wherein the ELISA plate comprises a solid phase carrier and a functional layer modified on the surface of the solid phase carrier, and the functional layer can generate stronger adsorption with protein biomolecules such as antigen or antibody and the like, so that the coating effect of the ELISA plate in the ELISA adsorption determination process can be improved.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, the invention provides an ELISA plate, which comprises a solid phase carrier and a functional layer modified on the surface of the solid phase carrier, wherein the functional layer comprises a graft copolymer, and a functional monomer of the graft copolymer comprises N-octadecyl acrylamide and/or N- (3-dimethylaminopropyl) methacrylamide.
In the ELISA plate, the functional layer contains hydrophobic groups or carries charges, the hydrophobic groups can generate adsorption with similar and compatible structure with the hydrophobic region of protein molecules, and the carried charges can generate electrostatic adsorption with the protein molecules carrying the charges, so that the functional layer can generate stronger adsorption with the protein molecules such as antigens or antibodies, and the effect of coating the protein molecules such as the antigens or the antibodies by the ELISA plate is improved.
Preferably, the solid support comprises polystyrene.
Preferably, the graft copolymer is a polymer formed by graft copolymerization of N-octadecyl acrylamide and N- (3-dimethylaminopropyl) methacrylamide.
In the invention, the hydrophobic group of the N-octadecyl acrylamide can generate stronger adsorption with the hydrophobic region of protein biomolecules, while the amino group of the N- (3-dimethylaminopropyl) methacrylamide can generate stronger electrostatic adsorption with the protein biomolecules in aqueous solution, and the amino group and the protein biomolecules are matched with each other to further enhance the adsorption of the ELISA plate on the proteins.
Preferably, the mass ratio of the N-octadecyl acrylamide to the N- (3-dimethylaminopropyl) methacrylamide is (0.5-1): 1, including but not limited to 0.6:1, 0.65:1, 0.7:1, 0.8:1, 0.9:1 or 0.95: 1.
In the invention, the adsorption effect of the ELISA plate on protein can be further enhanced by controlling the mass ratio of N-octadecyl acrylamide to N- (3-dimethylaminopropyl) methacrylamide to be (0.5-1): 1.
In a second aspect, the present invention provides a method for preparing the microplate according to the first aspect, the method comprising:
and cleaning and drying the solid phase carrier, putting the solid phase carrier into vacuum plasma processing equipment, vacuumizing, introducing process gas into the reaction chamber, adding a functional monomer, and performing vacuum plasma processing to obtain the elisa plate.
Preferably, the washing treatment includes washing with absolute ethanol and distilled water in this order.
Preferably, the cleaning process is performed in an ultrasonic cleaning machine.
Preferably, the washing time of the absolute ethyl alcohol is 8-12 min, including but not limited to 9min, 10min or 11 min.
Preferably, the cleaning time of the distilled water is 8-12 min, including but not limited to 9min, 10min or 11 min.
Preferably, the vacuum is pumped to 10-17 Pa, including but not limited to 11Pa, 12Pa, 13Pa, 15Pa or 16 Pa.
Preferably, the process gas comprises argon.
Preferably, the power of the vacuum plasma treatment is 60-80W, including but not limited to 62W, 65W, 68W, 70W, 72W, 75W or 78W.
Preferably, the time of the vacuum plasma treatment is 5-8 min, including but not limited to 6min, 6.5min, 7min or 7.5 min.
As a preferred technical scheme, the preparation method of the ELISA plate comprises the following steps:
(1) sequentially cleaning the solid phase carrier with absolute ethyl alcohol and distilled water in an ultrasonic cleaning machine for 8-12 min respectively, and drying at 40-45 ℃ for later use;
(2) putting the cleaned solid phase carrier into vacuum plasma processing equipment, starting a vacuum pump to vacuumize to 10-17 Pa, introducing argon into a reaction chamber, adding a functional monomer, starting the vacuum plasma processing equipment, setting the power to be 60-80W, and reacting for 5-8 min to obtain the ELISA plate.
In a third aspect, the present invention provides a kit comprising the microplate of the first aspect.
Preferably, the kit further comprises a sample diluent, a washing solution, a substrate developing solution and a reaction stopping solution.
In a fourth aspect, the invention provides the use of an elisa plate according to the first aspect and/or a kit according to the third aspect in an enzyme-linked immunosorbent assay.
Compared with the prior art, the invention has the following beneficial effects:
(1) according to the invention, the functional layer is modified on the solid phase carrier of the ELISA plate, so that the surface of the ELISA plate can generate a strong adsorption effect with protein molecules such as antigen or antibody, the effect of coating the protein molecules such as antigen or antibody by the ELISA plate is improved, and the sensitivity of ELISA test is improved;
(2) the protein saturation adsorption values of the ELISA plate are all larger than 342ng/cm2In addition, by simultaneously adopting two functional monomers of N-octadecyl acrylamide and N- (3-dimethylaminopropyl) methacrylamide and controlling the mass ratio of the two functional monomers to be (0.5-1): 1, the saturated adsorption value of the protein can be further improved, the internal CV can be reduced, and the saturated adsorption value of the protein of the enzyme label plate is higher than 515ng/cm2At most 761ng/cm2And the CV in the batch is lower than 4.7 percent, so that the adsorption capacity and the adsorption uniformity of the ELISA plate to the protein are further improved.
Detailed Description
For the purpose of facilitating an understanding of the present invention, the present invention will now be described by way of examples. It should be understood by those skilled in the art that the examples are only for the understanding of the present invention and should not be construed as the specific limitations of the present invention.
The examples do not show the specific techniques or conditions, according to the technical or conditions described in the literature in the field, or according to the product specifications. The reagents or apparatus used are conventional products commercially available from normal sources, not indicated by the manufacturer.
Example 1
The embodiment provides an ELISA plate, which comprises a solid phase carrier and a functional layer modified on the surface of the solid phase carrier, wherein the solid phase carrier is polystyrene, and the functional layer is a graft copolymer composed of N-octadecyl acrylamide. The preparation method of the ELISA plate comprises the following steps:
(1) sequentially washing the polystyrene plates with absolute ethyl alcohol and distilled water in an ultrasonic cleaning machine for 12min respectively, and drying at 40 ℃ for later use;
(2) putting the cleaned polystyrene plate between two parallel polar plates of low-temperature plasma processing equipment, wherein an orifice is upward, and closing the reaction chamber;
(3) starting a vacuum pump to vacuumize to 11Pa, opening a flow control valve, introducing argon (0.1L/min) and N-octadecyl acrylamide (0.34g/min) into a reaction chamber, setting the time to be 5min and the power to be 60w, and starting a generator;
(4) flushing, exhausting, opening the reaction chamber and sampling.
Example 2
The embodiment provides an ELISA plate, which comprises a solid phase carrier and a functional layer modified on the surface of the solid phase carrier, wherein the solid phase carrier is polystyrene, and the functional layer is a graft copolymer composed of N- (3-dimethylaminopropyl) methacrylamide. The preparation method of the ELISA plate comprises the following steps:
(1) sequentially washing the polystyrene plates with absolute ethyl alcohol and distilled water in an ultrasonic cleaning machine for 8min respectively, and drying at 45 ℃ for later use;
(2) putting the cleaned polystyrene plate between two parallel polar plates of low-temperature plasma processing equipment, wherein an orifice is upward, and closing the reaction chamber;
(3) starting a vacuum pump to vacuumize to 15Pa, opening a flow control valve, introducing argon (0.1L/min) and N- (3-dimethylaminopropyl) methacrylamide (0.37g/min) into a reaction chamber, setting the time to be 5min and the power to be 60w, and starting a generator;
(4) flushing, exhausting, opening the reaction chamber and sampling.
Example 3
The embodiment provides an ELISA plate, which comprises a solid phase carrier and a functional layer modified on the surface of the solid phase carrier, wherein the solid phase carrier is polystyrene, the functional layer is a graft copolymer composed of N-octadecyl acrylamide and N- (3-dimethylaminopropyl) methacrylamide, and the mass ratio of the N-octadecyl acrylamide to the N- (3-dimethylaminopropyl) methacrylamide is 0.5: 1. The preparation method of the ELISA plate comprises the following steps:
(1) sequentially washing the polystyrene plates with absolute ethyl alcohol and distilled water in an ultrasonic cleaning machine for 8min respectively, and drying at 42 ℃ for later use;
(2) putting the cleaned polystyrene plate between two parallel polar plates of low-temperature plasma processing equipment, wherein an orifice is upward, and closing the reaction chamber;
(3) starting a vacuum pump to vacuumize to 10Pa, opening a flow control valve, introducing argon (0.1L/min), N-octadecyl acrylamide (0.12g/min) and N- (3-dimethylaminopropyl) methacrylamide (0.24g/min) into a reaction chamber, setting the time to be 5min and the power to be 60w, and starting a generator;
(4) flushing, exhausting, opening the reaction chamber and sampling.
Example 4
The embodiment provides an ELISA plate, which comprises a solid phase carrier and a functional layer modified on the surface of the solid phase carrier, wherein the solid phase carrier is polystyrene, the functional layer is a graft copolymer composed of N-octadecyl acrylamide and N- (3-dimethylaminopropyl) methacrylamide, and the mass ratio of the N-octadecyl acrylamide to the N- (3-dimethylaminopropyl) methacrylamide is 0.8: 1. The preparation method of the ELISA plate comprises the following steps:
(1) sequentially washing the polystyrene plates with absolute ethyl alcohol and distilled water in an ultrasonic cleaning machine for 8min respectively, and drying at 43 ℃ for later use;
(2) putting the cleaned polystyrene plate between two parallel polar plates of low-temperature plasma processing equipment, wherein an orifice is upward, and closing the reaction chamber;
(3) starting a vacuum pump to vacuumize to 17Pa, opening a flow control valve, introducing argon (0.1L/min), N-octadecyl acrylamide (0.16g/min) and N- (3-dimethylaminopropyl) methacrylamide (0.2g/min) into a reaction chamber, setting the time to be 5min and the power to be 80w, and starting a generator;
(4) flushing, exhausting, opening the reaction chamber and sampling.
Example 5
The embodiment provides an ELISA plate, which comprises a solid phase carrier and a functional layer modified on the surface of the solid phase carrier, wherein the solid phase carrier is polystyrene, the functional layer is a graft copolymer composed of N-octadecyl acrylamide and N- (3-dimethylaminopropyl) methacrylamide, and the mass ratio of the N-octadecyl acrylamide to the N- (3-dimethylaminopropyl) methacrylamide is 1: 1. The preparation method of the ELISA plate comprises the following steps:
(1) sequentially washing the polystyrene plates with absolute ethyl alcohol and distilled water in an ultrasonic cleaning machine for 8min respectively, and drying at 45 ℃ for later use;
(2) putting the cleaned polystyrene plate between two parallel polar plates of low-temperature plasma processing equipment, wherein an orifice is upward, and closing the reaction chamber;
(3) starting a vacuum pump to vacuumize to 15Pa, opening a flow control valve, introducing argon (0.1L/min), N-octadecyl acrylamide (0.18g/min) and N- (3-dimethylaminopropyl) methacrylamide (0.18g/min) into a reaction chamber, setting the time to be 8min and the power to be 80w, and starting a generator;
(4) flushing, exhausting, opening the reaction chamber and sampling.
Example 6
The embodiment provides an ELISA plate, which comprises a solid phase carrier and a functional layer modified on the surface of the solid phase carrier, wherein the solid phase carrier is polystyrene, the functional layer is a graft copolymer composed of N-octadecyl acrylamide and N- (3-dimethylaminopropyl) methacrylamide, and the mass ratio of the N-octadecyl acrylamide to the N- (3-dimethylaminopropyl) methacrylamide is 0.3: 1. The preparation method of the ELISA plate comprises the following steps:
(1) sequentially washing the polystyrene plates with absolute ethyl alcohol and distilled water in an ultrasonic cleaning machine for 8min respectively, and drying at 45 ℃ for later use;
(2) putting the cleaned polystyrene plate between two parallel polar plates of low-temperature plasma processing equipment, wherein an orifice is upward, and closing the reaction chamber;
(3) starting a vacuum pump to vacuumize to 11Pa, opening a flow control valve, introducing argon (0.1L/min), N-octadecyl acrylamide (0.08g/min) and N- (3-dimethylaminopropyl) methacrylamide (0.27g/min) into a reaction chamber, setting the time to be 5min and the power to be 80w, and starting a generator;
(4) flushing, exhausting, opening the reaction chamber and sampling.
Example 7
The embodiment provides an ELISA plate, which comprises a solid phase carrier and a functional layer modified on the surface of the solid phase carrier, wherein the solid phase carrier is polystyrene, the functional layer is a graft copolymer composed of N-octadecyl acrylamide and N- (3-dimethylaminopropyl) methacrylamide, and the mass ratio of the N-octadecyl acrylamide to the N- (3-dimethylaminopropyl) methacrylamide is 1.2: 1. The preparation method of the ELISA plate comprises the following steps:
(1) sequentially washing the polystyrene plates with absolute ethyl alcohol and distilled water in an ultrasonic cleaning machine for 8min respectively, and drying at 40 ℃ for later use;
(2) putting the cleaned polystyrene plate between two parallel polar plates of low-temperature plasma processing equipment, wherein an orifice is upward, and closing the reaction chamber;
(3) starting a vacuum pump to vacuumize to 11Pa, opening a flow control valve, introducing argon (0.1L/min), N-octadecyl acrylamide (0.2g/min) and N- (3-dimethylaminopropyl) methacrylamide (0.16g/min) into a reaction chamber, setting the time to be 5min and the power to be 80w, and starting a generator;
(4) flushing, exhausting, opening the reaction chamber and sampling.
Test example 1 measurement of protein adsorption Capacity and adsorption uniformity of ELISA plate
This test example tested the protein adsorption capacity of the microplate prepared in examples 1-7.
The method comprises the following steps of taking an ELISA plate as a carrier, coating the ELISA plate by using normal human IgG (antibody) with the concentration of 50 mu g/mL, coating the ELISA plate overnight at 4 ℃, detecting the protein concentration of coating liquid before coating and liquid in holes of the ELISA plate after coating by using a BCA method trace protein concentration determination kit, calculating the difference value of the protein content of the liquid before and after coating, and judging the adsorption capacity of the ELISA plate, wherein the method specifically comprises the following steps:
1) preparing a standard curve solution;
2) calculating the OD of each sample of each point meter of the standard curve562Mean, mean minus OD of blank well (0 dots)562The average value is the calibrated OD of each point562；
3) Calibration OD with points of the Standard Curve562The value is the ordinate, the corresponding protein concentration is the abscissa, draw the standard curve on EXCEL, obtain the curvilinear equation;
4) then according to the curve equation and the calibrated OD of each sample562Calculating the original protein concentration of the coating liquid and the protein concentration of the liquid in the hole after each ELISA plate to be detected is coated overnight, wherein the concentration difference between the original protein concentration and the protein concentration is the protein saturated adsorption concentration (ng/mL) of each ELISA plate;
5) according to the structural size of the ELISA plate hole, the volume of the coating solution can be converted into the coating area (cm) in the ELISA plate hole2) Then the saturated adsorption value of the protein of the enzyme label plate is calculated as the protein adsorption mass divided by the coating surface area。
This test example tested the uniformity of protein adsorption of the microplate prepared in examples 1-7.
An enzyme label plate is used as a carrier, normal human IgG (antibody) with the concentration of 50 mu g/mL is used for coating, the enzyme label plate is coated overnight at the temperature of 4 ℃, an anti-human IgG enzyme conjugate is matched, the OD value of each hole is detected by double wavelengths (main wavelength 450nm and reference wavelength 630nm) through a color reaction, the light transmittance of each hole is calculated according to the OD value, and then the light transmittance mean value and the Coefficient of Variation (CV) are calculated, wherein the calculation method of the CV comprises the following steps:
1)OD＝OD450-OD630；
2) calculating an OD average value;
3) calculating OD standard deviation;
4) CV-standard deviation ÷ mean value.
The results are shown in tables 1 and 2.
TABLE 1
TABLE 2
As shown in tables 1 and 2, in both examples 1 and 2, the enzyme label plate is prepared by using a functional monomer (N-octadecyl acrylamide or N- (3-dimethylaminopropyl) methacrylamide), and the saturation adsorption value of the prepared enzyme label plate protein is higher than 342ng/cm2The maximum value is 381ng/cm2The Coefficient of Variation (CV) in batch is higher than 10.8%; in examples 3-7, two kinds of functional monomers (N-octadecylacrylamide and N- (3-dimethylaminopropyl) methacrylamide) were used to prepare an ELISA plate, and in examples 3-5, N-octadecylacrylamide and N- (3-dimethylaminopropyl) methylpropane were controlledThe mass ratio of the enamides is (0.5-1): 1, and the saturated adsorption value of the protein of the prepared ELISA plate is higher than 515ng/cm2At most 761ng/cm2The Coefficient of Variation (CV) in batches is lower than 4.7 percent, the mass ratio of the N-octadecyl acrylamide to the N- (3-dimethylaminopropyl) methacrylamide in examples 6 and 7 is not controlled to be (0.5-1): 1, and the protein saturation adsorption value of the prepared ELISA plate is higher than 411ng/cm2Maximum 489ng/cm2The Coefficient of Variation (CV) in the batch was less than 10.2% and at the lowest 6.5%.
Compared with the ELISA plates prepared in the embodiments 1 and 2, the ELISA plates prepared in the embodiments 3 to 7 have higher protein saturation adsorption value and lower intra-batch variation coefficient, which shows that the invention adopts N-octadecyl acrylamide and N- (3-dimethylaminopropyl) methacrylamide to effectively improve the protein saturation adsorption value of the ELISA plate and reduce the intra-batch variation coefficient, namely, the protein adsorption capacity and uniformity of the ELISA plate are improved; in addition, compared with the ELISA plates prepared in the embodiments 6 and 7, the ELISA plates prepared in the embodiments 3-5 have higher protein saturation adsorption value and lower intra-batch variation coefficient, which shows that the mass ratio of N-octadecyl acrylamide to N- (3-dimethylaminopropyl) methacrylamide is controlled to be (0.5-1): 1, so that the protein saturation adsorption value of the ELISA plate can be further improved, the intra-batch variation coefficient can be reduced, and the adsorption capacity and the adsorption uniformity of the ELISA plate on the protein can be further improved.
In conclusion, the functional layer is modified on the solid phase carrier of the enzyme label plate, so that the surface of the enzyme label plate can generate stronger adsorption with protein molecules such as antigen or antibody, the effect of coating the protein molecules such as antigen or antibody by the enzyme label plate is improved, and the adsorption capacity and the adsorption uniformity of the enzyme label plate to the protein are further improved by simultaneously adopting two functional monomers of N-octadecyl acrylamide and N- (3-dimethylaminopropyl) methacrylamide and controlling the mass ratio of the two functional monomers.
The applicant states that the present invention is illustrated by the above examples to show the detailed process equipment and process flow of the present invention, but the present invention is not limited to the above detailed process equipment and process flow, i.e. it does not mean that the present invention must rely on the above detailed process equipment and process flow to be implemented. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.
Claims (5)
1. An ELISA plate is characterized by comprising a solid phase carrier and a functional layer modified on the surface of the solid phase carrier;
the functional layer comprises a graft copolymer;
the graft copolymer is a polymer formed by graft copolymerization of N-octadecyl acrylamide and N- (3-dimethylaminopropyl) methacrylamide;
the mass ratio of the N-octadecyl acrylamide to the N- (3-dimethylaminopropyl) methacrylamide is (0.5-1.0) to 1;
the solid phase carrier is polystyrene.
2. A method for preparing an elisa plate according to claim 1, comprising:
sequentially cleaning the solid phase carrier with absolute ethyl alcohol and distilled water in an ultrasonic cleaning machine for 8-12 min respectively, drying at 40-45 ℃ for later use, putting the solid phase carrier into vacuum plasma treatment equipment, starting a vacuum pump to vacuumize to 10-17 Pa, introducing argon into a reaction chamber, adding N-octadecyl acrylamide and N- (3-dimethylaminopropyl) methacrylamide, starting the vacuum plasma treatment equipment, setting the power to be 60-80W, and reacting for 5-8 min to obtain the ELISA plate.
3. A kit comprising the microplate of claim 1.
4. The kit according to claim 3, further comprising a sample diluent, a washing solution, a substrate developing solution and a termination reaction solution.
5. Use of an elisa plate according to claim 1 and/or a kit according to claim 3 in an enzyme-linked immunosorbent assay.
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| CN101048222A (en) * | 2004-09-30 | 2007-10-03 | 麦克马斯特大学 | Composite material comprising layered hydrophilic coatings |
| CN106474946A (en) * | 2016-10-17 | 2017-03-08 | 东华大学 | A kind of preparation method of the polymeric film with hydrophilic antifouling property |
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