HK1081599B - Method for producing polymer layers - Google Patents
Method for producing polymer layers Download PDFInfo
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- HK1081599B HK1081599B HK06101610.2A HK06101610A HK1081599B HK 1081599 B HK1081599 B HK 1081599B HK 06101610 A HK06101610 A HK 06101610A HK 1081599 B HK1081599 B HK 1081599B
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- polymer coating
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- liquid composition
- indicator
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Description
The present invention relates to a method for producing a polymer coating on a support by photo-polymerization of a polymerizable liquid composition, to an apparatus suitable for producing a polymer coating on a support and to a method for producing a sensor comprising a polymer coating having an indicator incorporated therein.
The preparation of polymeric coatings on supports is a known technique. For this purpose, it is necessary to apply the polymerizable liquid to the support in as uniform a thickness as possible and to carry out complete polymerization. The polymerization process may be initiated by a chemical polymerization initiator or by irradiation of the liquid.
Polymerization techniques are known for the preparation of adhesive films, in which a mixture of functionalized acrylate prepolymers is applied to a support and then completely or partially hardened by irradiation of the surface of the polymer mass. Control of the polymer coating thickness can be achieved by selection of the intensity and duration of light exposure and by self-absorption of the prepolymer used.
A disadvantage of the prior art methods is that it is often very costly to adjust a predetermined and uniform polymer coating thickness on the carrier. Furthermore, in conventional polymerization processes, it is often found that the adhesion of the polymer coating to the support is very low.
The object of the present invention is therefore to overcome at least partially the disadvantages of the prior art described above, and in particular to provide a method for producing a polymer coating on a support, which method allows polymer coatings having a predetermined and uniform coating thickness to be produced in a relatively simple manner.
In contrast to the methods described in the prior art for preparing adhesives, in which a closed and hydrophobic polymer coating is prepared, hydrophilic coatings are prepared which are open and which absorb aqueous sample liquid and analytes.
This object is achieved by applying a liquid polymerizable composition to a support and carrying out the polymerization directly on the support, but not completely but only in a layer having a predetermined thickness. In this way, a coated carrier which adheres uniformly and well to the carrier can be produced simply.
The invention therefore relates to a method for producing a polymer coating on a transparent support, comprising the following steps:
(a) providing a carrier, and making the carrier into a shape,
(b) the photopolymerizable liquid composition is applied to a support,
(c) irradiating the photopolymerizable liquid composition through the support in such a manner that the liquid composition undergoes only partial polymerization and a polymer layer having a predetermined thickness is formed on the support, and
(d) the residual liquid composition is removed from the polymer coating.
The support used for the preparation of the polymer coating is an at least partially optically transparent support, for example a plastic support such as a polycarbonate film, a cellulose acetate film, a polyester film or a polyether film, a glass support or a quartz support. Supports made of composite materials may also be used. When such an at least partially optically transparent support is used, it is preferred that the liquid composition is irradiated through the support. The thickness of the support is preferably selected such that the polymer coating has sufficient mechanical stability on the one hand and sufficient transmission of the light used for irradiation on the other hand. For example, the thickness of the support used may be 5 μm to 20 mm. It is also preferred to use a UV transparent carrier.
The photopolymerizable liquid composition contains at least one photopolymerizable substance, i.e. a substance which is polymerizable by light, if necessary in the presence of a photoinitiator. Preferred examples of such substances are photopolymerizable monomers, such as ethylenically unsaturated substances, i.e. substances having a C ═ C bond. Further examples of suitable photopolymerizable substances are oligomers or polymers which are functionalized and can be crosslinked under light. Such photoactive functionalization products include, for example, acrylates, azides, carbazides, sulfone azides (sulfoazides), diazoketones, dimethylmaleimides, photocyclizable groups (e.g., chalcones), and Benophen derivatives. Suitable oligomers or polymers are, for example, polyurethanes, polyvinyl alcohols, polyesters, polyethers, polyvinyl pyrrolidones, polyacrylates or oligosaccharides. Particularly preferred are photopolymerizable substances selected from the group consisting of, for example, acrylic monomers, such as acrylamide, acrylic esters, such as polyethylene glycol diacrylate, vinyl aromatic monomers, such as 4-vinylbenzenesulfonic acid, functionalized polyvinylpyrrolidone and any combination of one or more of the foregoing.
In a preferred embodiment, aqueous polymerizable compositions are used in which the monomers to be polymerized are present in dissolved form. Therefore, it is desirable for this embodiment to use a hydrophilic monomer having sufficiently high solubility in an aqueous solvent.
Initiation of the polymerization reaction in the liquid composition may be effected by irradiation with light. To initiate the polymerization reaction, it is preferred that the composition contains one or more photoinitiators. Examples of suitable photoinitiators are free radical initiators such as benzophenones, benzidines, anthraquinones, thiosulfonic acids, azo compounds or ionic initiators such as triarylsulfonium salts, arylsulfonium hexafluoroantimonates.
The composition is illuminated through the carrier so that the polymerization starts at the carrier surface and-depending on the set polymerization conditions-the polymerization ends at a predetermined distance from the carrier surface due to the absorption of light into the polymerizable liquid. The warping or deviation of the horizontal layer on the support does not affect the thickness of the polymer coating. Mechanical tolerances can be further eliminated.
The thickness of the polymer coating can be adjusted within wide limits by suitable measures. In particular the thickness of the polymer coating can be controlled by varying the intensity of the light irradiation, the duration of the light irradiation or/and the addition of polymerization inhibitors, for example UV-absorbing substances, to the liquid composition. In addition, control can also be effected by the thickness of the support or/and the support material. The thickness of the polymer coating is preferably & lt, 500 μm and particularly preferably & lt, 100. mu.m. If the polymer coating is used as a sensor, in particular as a biosensor, it is often also possible to produce smaller layer thicknesses, preferably & lt, 50 μm, particularly preferably & lt, 5 μm.
If a polymer coating is used as a component of the sensor, it should preferably comprise, in addition to the above-mentioned components, indicators, such as optical and/or electrochemical indicators, which are responsive to the parameter in the medium surrounding the polymer layer. The photopolymerizable liquid compositions can be added to the indicators or, especially when they are small molecules, the indicators can be dispersed into the finished polymer coating. If the polymer layer contains multiple indicators, a combination of the above measures is also possible.
The indicator may also be a macromolecule, for example having a molecular weight of 10kD or more, especially 20kD or more. Particularly preferred for use as an indicator is a catalyst material, for example an enzyme which may, if desired, be present in the form of an enzyme-coenzyme complex. Examples of particularly preferred enzymes are oxidoreductases, in particular dehydrogenases, such as glucose dehydrogenase (e.c.1.1.1.47) or oxidases. The coenzyme is preferably an organic molecule which is covalently or non-covalently linked to the enzyme and which is capable of being denatured, e.g.oxidized or reduced, by reaction with the enzyme substrate. Examples of preferred coenzymes are flavin derivatives, nicotine derivatives, quinone derivatives, e.g. FAD, FADH2，NAD+，NADH/H+，NADP+，NADPH/H+Or PQQ.
In addition to the enzyme and, if appropriate, the coenzyme, the polymer layer also contains a mediator, i.e.a substance which has the effect of regenerating the coenzyme. In this case the enzyme acts as a catalytic indicator, i.e. it is capable of converting a plurality of molecules of a substrate, e.g. an analyte present in the sample to be added, such as glucose in blood.
In a particularly preferred embodiment, the polymeric coating contains an enzyme-coenzyme complex as a stoichiometric reactant for the enzyme substrate to be validated. The coenzyme is not regenerated in one reaction. In this embodiment, no further mediator is necessary which is required in combination with the complex mixture of reactants which is less stable and highly susceptible to interference.
If a macromolecular substrate is used as the indicator, a crosslinked polymeric coating may be obtained by suitable polymer crosslinking (e.g., by using di-or/and polyfunctional monomers) in which the macromolecular indicator substance is incorporated in immobilized form, while low molecular weight substances, such as coenzymes, enzyme substrates, and the like, may be dispersed in the layer.
The preparation of the polymer coating may be carried out in a continuous process, wherein the polymer coating is continuously produced on the liquid photopolymerizable composition applied to the support. When the polymerization is carried out continuously, the photopolymerizable liquid composition is preferably applied continuously to the self-moving support at a first location and then continuously illuminated at a second location. Obviously, the carrier may also be held stationary while the location of the applied liquid and the light is moved. It is also contemplated that a discontinuous process may be used to produce the polymer film. These embodiments have in common that, due to the irradiation of the photopolymerizable liquid composition through the support, a liquid polymerization reaction can be carried out which starts directly on the support but which is not complete.
A further subject of the invention is an apparatus for producing a polymer coating, comprising
(a) Means for receiving and if necessary also for transporting the carrier,
(b) a mechanism for applying a photopolymerizable liquid composition to a support, and
(c) means for irradiating the photopolymerizable liquid composition through the support to thereby form a polymer coating having a predetermined thickness on the support, and
(d) optionally, a mechanism for separating the unpolymerized liquid composition from the polymer coating.
The method and apparatus can be used to produce a sensor in which an indicator, for example a biomolecule such as an enzyme, is embedded in a polymeric coating. The indicator may be present in immobilized form in the polymeric coating. Particularly preferred indicators are enzymes, if necessary in the form of enzyme-coenzyme complexes. The sensor may be, for example, an optical or/and electrochemical sensor. Particularly preferred sensors are fluorescence based sensors.
Yet another aspect of the invention is a method of making a sensor, comprising the steps of:
(a) preparing a carrier, namely preparing a carrier,
(b) applying a photopolymerizable liquid composition comprising at least one indicator to the support,
(c) irradiating the photopolymerizable liquid composition through the support in such a manner as to form a polymer coating having a predetermined thickness on the support,
(d) removing residual liquid composition from the polymer coating, and
(e) the carrier with the polymer coating containing the indicator is placed in a sensor that contains a mechanism for detecting the reaction between the indicator and the analyte in the sample.
The detection means are preferably optical or/and electrochemical detection means. Particularly preferred is again an optical detection means comprising a light source to illuminate the polymer coating and a detector to capture light from the polymer coating. The light source, such as a laser or LED, is preferably arranged to shine light through the carrier into the polymer coating. The detector is preferably arranged to capture light, e.g. fluorescent emissions, from the polymer coating.
The sensor may be used to determine any analyte, e.g. a physicochemical parameter, e.g. temperature, e.g. O2、CO2、NOxEtc. or for determining biochemical parameters, e.g. analyzed in biological samplesAn object, such as a body fluid.
In addition, the invention will be elucidated below by means of figures and examples.
Fig. 1 shows a first embodiment of a sensor prepared by the method of the present invention. An optically transparent support (1) is coated with a polymer coating (2) containing an indicator, for example a detection reagent for enzymatic reactions. A sample (3), such as blood, is then placed on the polymer coating. The enzymatic reaction that takes place between the analyte contained in the sample (3) and the detection reagent contained in the polymer coating (2) can be determined by optical methods. Light from a light source (4), such as a laser or LED, is injected into the reagent layer (2) from below (through the carrier). The absorbed light or fluorescence reflected from the sample can be detected in a detector (5). If necessary (in particular for detecting fluorescence), an optical filter (6) is inserted upstream of the detector in order to suppress crosstalk of the fluorescence excitation light.
FIG. 2 shows the preparation of a polymer coating according to the invention. In a first position (13), for example on an optically transparent carrier (11), for example on a plastic film, liquid reagents (12) are applied. In the second position, the liquid reagent (12) is irradiated from below through the carrier (11) with light from the light source (14). While the carrier is moved in the direction (15) indicated by the arrow. The polymerized reagent layer (16) is then formed directly on the carrier (11). An excess of liquid reagent is present on the polymer coating (16). The thickness of the polymerized reagent layer (16) can be controlled by the reagent composition, the duration and intensity of the light exposure and by the properties of the support (11).
Fig. 3 shows an embodiment of the fluorescence-based sensor from below. For example, a polymer coating made by the continuous process of fig. 2 and containing an indicator may be cut and embedded into the sensor 21 using known techniques. After the sample is applied to the upper side, it is irradiated from below with excitation light (23) from a light source, for example, UV light. Fluorescence (24), such as blue light, produced by the reaction of the analyte with the detection reagent in the polymer coating (22) can be detected by a detector.
Multiple (the same or different) reactants may also be applied to the support. An example of such an embodiment is shown in fig. 4 as a disc. A plurality of reagent dots (32) consisting of a polymer coating with an indicator can be arranged on an optically transparent carrier (31).
Examples
Example 1: detection of glucose dehydrogenase System (GlucDH) NAD in Polymer membranes+Glucose in (1)
A suspension of the following was mixed into a plastic reagent bottle.
Formulation 1
| Substance(s) | Amount [ g ]] | Percent by weight [% ]] |
| Acrylamide | 2.5 | 22.02 |
| Methylene bisacrylamide | 0.7 | 6.17 |
| 2, 2-dimethoxy-2-phenylacetophenone | 0.05 | 0.44 |
| Glycerol | 5 | 44.05 |
| Hydroxyethyl methacrylate | 1.4 | 12.33 |
| Methacrylic acid methyl ester | 0.4 | 3.52 |
| Crodasinic O solution, pH8, 0.3g/1000ml | 1 | 8.81 |
| N, N' - (1, 2-dihydroxyethylene) bisacrylamide | 0.3 | 2.64 |
| Sum of | 11.35 | 100 |
0.5ml of this suspension and 0.5ml of glucDH solution (100mg/ml) were mixed, and the mixture was homogeneously mixed without bubbles in an ultrasonic bath.
The clear solution was cast onto corona-treated film (. The membrane was then washed briefly with water and then air dried.
The layer thickness thus obtained was < 2 μm. Freshly prepared glucose/NAD+Solution (GKL-3-solution, 300mg/d1 glucose, 1ml/6.4mg NAD+) Spotted on the membrane. Then a strong fluorescence was immediately observed under a UV lamp.
Example 2: influencing the layer thickness by adding UV absorbers
A polymer coating (formulation 2) was prepared which contained a blue pigment (absorption maximum ≈ 650nm) for better recognition. In another test, a yellow pigment was mixed into the initial formulation as a UV absorber (formulation 3).
Formulation 2
| Substance(s) | Measurement of | Percent by weight [% ]] |
| Acrylamide | 37.5g(0.53mol) | 25.78 |
| Poly (ethylene glycol diacrylate), Mw ≈ 575g/mol | 52.5g (about 0.96mol) | 36.10 |
| Solution of Crodasinic O (0.3g/ll) | 50g | 34.38 |
| 4-Vinylbenzenesulfonic acid | 5g | 3.44 |
| Photoinitiator 2, 2-dimethoxy-2-phenylacetophenone | 350mg | 0.24 |
| New Process for preparing methylene blue N | 100mg | 0.06 |
| Sum of | 145.45g | 100 |
The mixture was mixed well by stirring and ultrasonic bath treatment, dispersed on 140 μm Pokalon film (plasma treatment, grade 4) with a dropper and irradiated for 1min on a UV irradiator (ActinaU4, W.Lemmen GmbH).
The resulting layer thickness was measured with a micrometer screw and was 240.5. mu.m.
Formulation 3
| Substance(s) | Measurement of | Percent by weight [% ]] |
| Formulation 2 | 1ml | About 99.99 |
| Mordant yellow 7(686 number) (UV absorber) | 0.0001g | 0.001 |
| Total amount of | About 1.0001g | 100 |
The mixture as described above is distributed on a membrane and then polymerized. The resulting layer thickness was 79.3 μm as measured with a micrometer screw.
Experimental results show that an effect on layer thickness can be achieved. If no UV absorber is present, the layer thickness is 240.5 μm under otherwise identical reaction conditions (see above); whereas with the UV absorber (mordant yellow 7) only 79.3 μm.
Claims (33)
1. A method of preparing a polymeric coating on a transparent support comprising the steps of:
(a) preparing a carrier, namely preparing a carrier,
(b) the photopolymerizable liquid composition is applied to a support,
(c) irradiating the photopolymerizable liquid composition through the support in such a manner as to form a polymer coating having a predetermined thickness on the support, and
(d) the residual liquid composition is removed from the polymer coating.
2. The method of claim 1, characterized in that an at least partially optically transparent support is used.
3. The process as claimed in claim 1, characterized in that a support having a thickness of at least 5 μm is used.
4. The process as claimed in claim 1, characterized in that a support selected from the group consisting of plastic supports, glass supports or quartz supports is used.
5. A process according to claim 1, characterised in that a photopolymerizable liquid composition is used which comprises at least one photopolymerizable substance.
6. A process according to claim 5, characterized in that the photopolymerizable substance used is selected from the group consisting of acrylic monomers, vinyl aromatic monomers, functionalized polyvinylpyrrolidone and any combination thereof.
7. A process according to claim 1, characterized in that the photopolymerizable liquid composition used contains at least one photoinitiator.
8. The process of claim 1, characterized in that the polymer coating thickness is controlled by varying three variables:
(i) the intensity of the irradiation is varied according to the intensity of the irradiation,
(ii) duration of irradiation, and/or
(iii) A substance capable of absorbing the light of the polymerization reaction is added.
9. A process according to claim 1, characterized in that the thickness of the polymer coating produced is 500 μm or less.
10. The process as claimed in claim 9, wherein the polymer coating produced has a thickness of 5 μm or less.
11. A method according to claim 1, characterized in that the polymer coating produced contains at least one indicator.
12. A method according to claim 11, characterized in that a macromolecular substance is used as indicator.
13. The method according to claim 11, characterized in that a catalytic substance is used as indicator.
14. The method of claim 13, characterized in that the catalytic species is an enzyme.
15. The method of claim 1, characterized in that a crosslinked polymer coating is prepared.
16. A method according to claim 15, characterized in that the crosslinked polymer coating comprises macromolecules.
17. The method of claim 1, characterized in that the irradiation is carried out with UV light.
18. The process as claimed in claim 1, wherein the polymerization is carried out as a continuous process.
19. The method of claim 18, characterized in that the photopolymerizable liquid composition is continuously applied to the self-moving support at a first location and then continuously illuminated at a second location.
20. An apparatus for producing a polymer coating, comprising:
(a) means for receiving and if necessary also for transporting the carrier,
(b) a mechanism for applying a photopolymerizable liquid composition to a support,
(c) means for irradiating the photopolymerizable liquid composition through the support to thereby form a polymer coating having a predetermined thickness on the support.
21. The device of claim 20, further comprising
(d) A mechanism for removing unpolymerized composition from the polymer coating.
22. Use of the method according to one of claims 1 to 19 or the device according to claim 20 or 21 for producing a sensor in which the polymer coating contains at least one indicator.
23. Use according to claim 22, characterized in that the indicator is present in immobilized form in the polymer coating.
24. Use according to claim 22 or 23, characterized in that the polymeric coating contains an enzyme.
25. Use according to claim 24, characterized in that the enzyme contained in the polymer coating is present in the form of an enzyme-coenzyme complex.
26. Use according to claim 22 or 23, characterized in that the sensor is selected from optical or/and electrochemical sensors.
27. A method for making a sensor, comprising the steps of:
(a) preparing a transparent carrier, and preparing a transparent carrier,
(b) applying a photopolymerizable liquid composition comprising at least one indicator to a support,
(c) irradiating the photopolymerizable liquid composition through the support in such a manner as to form a polymer coating having a predetermined thickness on the support,
(d) the residual liquid composition is removed from the polymer coating,
(e) the carrier with the indicator-containing polymer coating is placed in a sensor that contains a mechanism for detecting the reaction of the indicator and the analyte in the sample.
28. The method of claim 27, wherein the indicator comprises an enzyme.
29. The method of claim 28, characterized in that the indicator comprises an enzyme in the form of an enzyme-coenzyme complex.
30. The method of claim 28, characterized in that the detection means comprise optical or/and electrochemical detection means.
31. The method of claim 30, wherein the optical detection mechanism comprises a light source for illuminating the polymer coating and a detector for capturing light from the polymer coating.
32. The method of claim 31, wherein the light source is positioned so that light can be directed through the carrier and into the polymer coating.
33. The method of claim 31 or 32, characterized in that a detector is arranged to capture the fluorescence emission from the polymer coating.
Applications Claiming Priority (7)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE10221840.4 | 2002-05-16 | ||
| DE10221845.5 | 2002-05-16 | ||
| DE10221846.3 | 2002-05-16 | ||
| DE2002121840 DE10221840A1 (en) | 2002-05-16 | 2002-05-16 | Production of polymer layers on a transparent support, for use in sensors, e.g. for blood analysis, comprises coating the support with a photopolymerizable liquid composition and irradiating the liquid through the support |
| DE2002121845 DE10221845A1 (en) | 2002-05-16 | 2002-05-16 | Detecting analyte by enzymatic reaction, useful specifically for measuring glucose in blood, based on reaction with enzyme-coenzyme complex |
| DE2002121846 DE10221846A1 (en) | 2002-05-16 | 2002-05-16 | Detecting an analyte by enzymatic reaction, useful specifically for measuring glucose in blood, based on reaction with coenzyme and inactive coenzyme-binding protein |
| PCT/EP2003/005179 WO2003097859A2 (en) | 2002-05-16 | 2003-05-16 | Method for producing polymer layers |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| HK1081599A1 HK1081599A1 (en) | 2006-05-19 |
| HK1081599B true HK1081599B (en) | 2011-09-02 |
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