CA2131007A1 - One-way reaction vessel for the solid-phase immunological analysis of, and a method of measuring constituents which can be determined via immune reactions - Google Patents
One-way reaction vessel for the solid-phase immunological analysis of, and a method of measuring constituents which can be determined via immune reactionsInfo
- Publication number
- CA2131007A1 CA2131007A1 CA002131007A CA2131007A CA2131007A1 CA 2131007 A1 CA2131007 A1 CA 2131007A1 CA 002131007 A CA002131007 A CA 002131007A CA 2131007 A CA2131007 A CA 2131007A CA 2131007 A1 CA2131007 A1 CA 2131007A1
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- Prior art keywords
- reaction vessel
- disposable reaction
- substrate
- vessel according
- determined
- Prior art date
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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
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/60—Construction of the column
- G01N30/6091—Cartridges
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/60—Construction of the column
- G01N30/6052—Construction of the column body
-
- 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
- G01N33/54366—Apparatus specially adapted for solid-phase testing
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/60—Construction of the column
- G01N30/6034—Construction of the column joining multiple columns
- G01N30/6039—Construction of the column joining multiple columns in series
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- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Hematology (AREA)
- Molecular Biology (AREA)
- General Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Pathology (AREA)
- Biomedical Technology (AREA)
- Physics & Mathematics (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Clinical Laboratory Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Automatic Analysis And Handling Materials Therefor (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
One-Way Reaction Vessel for the Solid-Phase Immunological Analysis of, and a Method of Measuring Constituents which can be determined via Immune Reactions Abstract The present invention concerns a one-way reaction vessel, open at both ends, for solid phase immunological analysis, with at least one immunologically reactive constituent applied onto a substrate with an active or activated surface. Said reactive constituent has a load density such that the constituent to be determined binds quantitatively (bonding equilibrium) to the reactive constituent before the flowing liquid leaves the substrate; the volume of the substrate bed is up to 600 ul, and the flow speed of the liquid from the substrate bed is determined accordingly. The one-way reaction vessel can be used to measure constituents which can be determined on the basis of immune reactions.
Description
- 1 - 2~3~0~' One-Way Reaction Vessel for the Solid-Phase Immunological ~nalysis of, and a Method of Measuring Constituents which can be determined via Immune Reactions Description The present invention concerns a disposable reaction vessel for the solid-phase immune analysis, and a method for its application.
Qffinity chromatography is a technology which is broadly applied in preparative purification of bio-molecules. Therein, advantage is taken of a specific interaction between the molecule to be determined and a complementary binding partner. In the general practice of such methods a sample containing the bio-molecule to be purified is applied to a chromatographic column, which contains one of' the mutually complementary binding partners bound to a solid substrate. Examples of pairs of complementary binding partners are enzymes and their substrates, antibodies and hapten resp. antigen, and mutually complementary DN~ or RNQ single chains.
In immunoaffinity chromatography, for quantification use is made of the interaction between an antigen resp. a hapten and the complementary antibody.
Up to now, in immunoassays using affinity chromatography reusable columns are generally used which are packed with the antibody or antigen (named ligand in the following) bound to a solid substrate. The sample to be analyzed is pressed through the chromatographic column by high pressure to guarantee a rapid chromatographic processing. The application of high-pressure liquid chromatography (HPLO) for immunoassays is described e.g. by de Elvis, W.U., and Wilson, G.S., in "~nalytical Chemistry", 1985, Vol. 57, pp. 2754-2756, and Sportsman7 3.R. et al. in "Qnalytical Chemistry", 1983, Vol. ~5, pp. 771-775. However, the use of HPLC
for immunoassays has several drawbacks On the one hand, the use of costly instruments is necessary for this technique, and on the other hand, the chromatographic columns for the analysis must meet high technical demands. The columns must withstand the high pressures applied in HPLC as well as be reusable to allow a rational application of this technology. However, the reuse of the columns re~uires additional measures before a new sample can be analyzed on the same column. ~s pubtished in DE-OS 37 11 894, reusable columns are regenerated prior to every analysis to prevent effects of displaced and incompletely eluted substances of the previrus test.
,. .,, ` ` . ,, , ,., ",, ~ i , . ' i ~ ;' i ' ~ i Z~ 3~007 ~ - 2 In the DE-OS 24 48 411 a reaction vessel is described for solid-phase immune analysis However, prior to a quantitative determination, this vessel for multiple use requires a calibration with standards of the component to be determined. Besides, long incubation times are necessary in this described reaction vessel, during which the flow through the column is stopped. 6 hours to 2 days are recommended to guarantee sufficient binding of the component to be determined to the immunologically complementarY
binding partner loaded onto the substrate However, the binding reaction is always terminated before the binding equilibrium is reached, which would take 1-2 days, to guarantee a rational performance The requirement of calibration with standard solutions is a consequence as we~l of this early termination of the binding reaction.
Rccordihg to the preference in practice for disposable columns, the DE-PS 26 26 8~3 describes a procedure for the preparation of a prefabricated disposable adsorption column especially for radio immunoassays using analytical instruments applying centrifugal forces. However, this column is not a reaction column, and is useful for the separation a~ter an immune reaction only, because it is not loaded with a specific immunoreactive component. To prevent disturbing effects during the separation via gel bed, the flow resistance of the porous retaining dèvices is kept as low as possible.
The object of the present invention is to provide a disposable reaction vessel for the solid-phase immune analysis which allows the rapid direct performance of immunoassays, which is easy to handle, and which is ready to immediate use without prior calibration or regeneration, and a method for the application of this vessel.
With respect to the device, this is accomplished in the present ,~ r invention by a disposable reaction vessel for the solid-phase immune analysis which is open at the upper and lower ends, in which at least one immunologically reactive component is bound to a substrate, characterized by a substrate bed volume of up to 600 ul, and by the determination of the liquid flow velocity by the permeability of the substrate. The substrate is either a solid frit, membran or the like, or it is contained between an upper and a lower separating device.
With respect to the method, this is accomplished by a procedure for the determination of components which can be determined Vi3 immune reactions, which is characterized by the application of the sample to be analyzed to d disposable reaction vessel according to any one of the claims 1-20, which is further characterized by the retention of the sample component, which has to be determined, by the complementary immunologically reactive component in the disposable reaction vessel during the direct flow-through, and in which the component to be determined is 2~3~00~
a) eluted and determined, or b) marked in the disposable reaction vessel via amplification reactions with common immunological markers, and then determined, or c) determined via a marked compound in reactive competition to the component to be determined.
The invention is described in detail with relation to figures 1 and 2 which show:
Fig. la a longitudinal section of a disposable reaction vessel 1 according to the invention containing two separating devices 2a and 2b, between which the substrate with the ' immunologically reactive component 3 is located, Fig. lb a top view of the disposable reaction vessel 1, and Fig. 2 a scheme of an online-system for antibody production control.
The dimensions given in figures la and lb are for example only.
The described disposable reaction vessel 1 and the described method, in which the disposable reaction vessel 1 is used, exhibit several considerable advantages compared to known reaction vessels resp. methods.
The described disposable reaction vessel 1 allows a rapid performance of the quantitative determination of components which can be determined by immunoreactions. The handling of the reaction vessel 1 is very simple and does not require the use of costly instruments such as HPLC apparatus. Besides excellent results are obtained with the aforesaid disposable reaction vessel 1 at extremely low consumption of substrate 3 with bound immunologically reactive component (named substrate bed in the following). R disposable reaction vessel 1 with a substrate bed volume 8, i.e. the volume between the upper and lower separating device 2a and 2b, of 50 ul is preferably used. The small substrate bed volume allows the use of small quantities of elution liquid with the consequence that the dilutio.- effect of the elution remains small, and thus the detection limit is not decreased in contrast to other methods in which larger elution volumes are necessary.
i`-'3` ~
Z131~0~7 Besides, the aforesaid reaction vessel 1 allows the quantitative determination at binding equilibrium (end-point determination) between the component to be determined and the immunologically reactive component. The binding equilibrium is reached within an incubation time of 1 minute already in the aforesaid disposable reaction vessel 1.
The rapid e~uilibration is rendered possible by the small substrate bed volume 8, its flow resistance, and the high loading of the substrate with the immunologically reactive component. The consequence of the controlled flow and the high loading of the substrate with the reactive component is that the component to be determined is bound quannatitatively to the reactive component within about 1 minute, i.e. before the flowing liqu;d leaves the substra,te bed volume. Therefore, a comparative analysis with standard references is spared.
Besides, a rapid analysis of samples is possible, because the disposable reaction vessel 1 allows immediate use, because the substrate 3 with the immunologically reactive component can be standardi~ed and ready-made, and the disposable reaction vessel 1 does not need to be calibrated before use.
The substrate 3 does not need to be stable against pressure, because the disposable reaction vessel 1 allows the rapid analysis of samples without application of HPLC. For the substrate 3, all common materials useful for affinity chromatography can be considered. Prefered substrate materials are polymeric sugars, plastics, plastic-modified substrates, metal oxides, or silicates.
Substrate materials in the form of frits are especially prefered.
For the determination of very large components such as viruses, which do not have access to the pores of such substrate materials, graft copolymers with side chains with active groups are prefered.
The immunologically reactive component loaded onto the substrate material 3 can bs bound to the substrate material covalently as well as adsorptively. The immunologically reactive component can be selected from the group containing haptens, antigens, antibodies, and immuno-affine proteins. Polyclonal as well as monoclonal antibodies can be used.
.. . . .
The vessel material 4 does not need to be stable against pressure, because the disposable reaction vessel 1 does not need to be subjected to high pressures in the present procedure. Therefore, besides pressure-resistant materials such as metals, other not pressure-resistant materials can be considered for the production of the disposable reaction vessel 1 such as glass, natural materials, and plastics. Prefered plastics are polyethylene~
polypropylene, and/or polystyrene, and prefered metals are aluminum and stainless steel.
For practi~cal purposes, the disposable reaction vessel 1 has a swelling 5 at one end to allow adaption of the reaction vessel into a holding and moving device for automatic transport. This is necessary for the use in automated sample processing devices.
~13~1007 ` - 5 -Besides, for practical purposes, the disposable reaction vessel 1 has a smaller-diameter joininy at the end opposite to the swelling, preferentially a joining ~or a socket-switch connection with another disposable reaction vessel~ This facilitates e.g.
joining in series of several disposable reaction vessels 1, whereby the outlet 6 of the first vessel is,connected with the inlet of the second vessel by a easy simple male-female connection.
Besides, the disposable reaction vessel 1 is closable by caps 7 and 9.
The present procedùre using said disposable reaction vessel 1 is superior due to, among others, a strongly simplified procedure compared to common procedures, and because it allows a rapid quantitative and qualitative determination. This is enabled partly because the determination of the component to be determined does not require a preceding calibration or regeneration of the used substrate in the disposable reaction vessel 1, and because no series reference measurements with standard solutions are ', necessary. The easy handling is a consequence of the possibility to apply samples and other solutions by usual laboratory equipment or by a pipetting automate. ~-Besides manual procedures, automated procedures are improved by application of the invented reaction vessel 1. In such procedures, the application of the sample and the further steps of the procedure ars performed by an automated sample processing device.
Such a sample processing device is used for example in the online production process control of antibodies produced in a fermenter.
In such procedures the samples are automatically applied in time intervalls of about 6 to 8 hours, and the antibody concentration in the fermentation medium can be continuously determined rapidly and easily over a long time period using the disposable reaction vessel 1. Q system for antibody production control using the invented disposable reaction vessel 1 is schematically described ~ ;
in fig. 2. Therein, the disposàble reaction vessel 1 is included in the automated sample preocessing device.
The present procedure can be performed rapidly without application of high pressures to the disposable reaction vessel 1 after sample application. But low suction or pressure forces can be used for quicker performance, e.g. by centrifugation or pumping.
If necessary, several disposable reaction vessels 1 can be connected in series or parallel in the present procedure. This allows e.g. the simultaneous determination of several parameters in one single sample resp. the simultaneous analysis of several samples.
.
2~3~007 The connection of several disposable reaction vessels in series is advantageous e.g. for the performance of allergy tests. Therein, a different antigen (allergen) is applied to each different column, then the columns are connected in series, and the sample containing the antibody of the class IgE which mediates the allergic reaction is applied to the first reaction vessel.
The sample flows successively through the different disposable reaction vessels 1 containing the different allergens. The IgE of the sample will be bound in the reaction vessel which contains the allergy-evoking antigen, and can be detected e.g. via fluorescence detection. In this way, one single sample can be tested simultaneously with many different potential allergens.
~ '''.;''' '` ~''''', ' ''' ; ' , ~ ' ;
Qffinity chromatography is a technology which is broadly applied in preparative purification of bio-molecules. Therein, advantage is taken of a specific interaction between the molecule to be determined and a complementary binding partner. In the general practice of such methods a sample containing the bio-molecule to be purified is applied to a chromatographic column, which contains one of' the mutually complementary binding partners bound to a solid substrate. Examples of pairs of complementary binding partners are enzymes and their substrates, antibodies and hapten resp. antigen, and mutually complementary DN~ or RNQ single chains.
In immunoaffinity chromatography, for quantification use is made of the interaction between an antigen resp. a hapten and the complementary antibody.
Up to now, in immunoassays using affinity chromatography reusable columns are generally used which are packed with the antibody or antigen (named ligand in the following) bound to a solid substrate. The sample to be analyzed is pressed through the chromatographic column by high pressure to guarantee a rapid chromatographic processing. The application of high-pressure liquid chromatography (HPLO) for immunoassays is described e.g. by de Elvis, W.U., and Wilson, G.S., in "~nalytical Chemistry", 1985, Vol. 57, pp. 2754-2756, and Sportsman7 3.R. et al. in "Qnalytical Chemistry", 1983, Vol. ~5, pp. 771-775. However, the use of HPLC
for immunoassays has several drawbacks On the one hand, the use of costly instruments is necessary for this technique, and on the other hand, the chromatographic columns for the analysis must meet high technical demands. The columns must withstand the high pressures applied in HPLC as well as be reusable to allow a rational application of this technology. However, the reuse of the columns re~uires additional measures before a new sample can be analyzed on the same column. ~s pubtished in DE-OS 37 11 894, reusable columns are regenerated prior to every analysis to prevent effects of displaced and incompletely eluted substances of the previrus test.
,. .,, ` ` . ,, , ,., ",, ~ i , . ' i ~ ;' i ' ~ i Z~ 3~007 ~ - 2 In the DE-OS 24 48 411 a reaction vessel is described for solid-phase immune analysis However, prior to a quantitative determination, this vessel for multiple use requires a calibration with standards of the component to be determined. Besides, long incubation times are necessary in this described reaction vessel, during which the flow through the column is stopped. 6 hours to 2 days are recommended to guarantee sufficient binding of the component to be determined to the immunologically complementarY
binding partner loaded onto the substrate However, the binding reaction is always terminated before the binding equilibrium is reached, which would take 1-2 days, to guarantee a rational performance The requirement of calibration with standard solutions is a consequence as we~l of this early termination of the binding reaction.
Rccordihg to the preference in practice for disposable columns, the DE-PS 26 26 8~3 describes a procedure for the preparation of a prefabricated disposable adsorption column especially for radio immunoassays using analytical instruments applying centrifugal forces. However, this column is not a reaction column, and is useful for the separation a~ter an immune reaction only, because it is not loaded with a specific immunoreactive component. To prevent disturbing effects during the separation via gel bed, the flow resistance of the porous retaining dèvices is kept as low as possible.
The object of the present invention is to provide a disposable reaction vessel for the solid-phase immune analysis which allows the rapid direct performance of immunoassays, which is easy to handle, and which is ready to immediate use without prior calibration or regeneration, and a method for the application of this vessel.
With respect to the device, this is accomplished in the present ,~ r invention by a disposable reaction vessel for the solid-phase immune analysis which is open at the upper and lower ends, in which at least one immunologically reactive component is bound to a substrate, characterized by a substrate bed volume of up to 600 ul, and by the determination of the liquid flow velocity by the permeability of the substrate. The substrate is either a solid frit, membran or the like, or it is contained between an upper and a lower separating device.
With respect to the method, this is accomplished by a procedure for the determination of components which can be determined Vi3 immune reactions, which is characterized by the application of the sample to be analyzed to d disposable reaction vessel according to any one of the claims 1-20, which is further characterized by the retention of the sample component, which has to be determined, by the complementary immunologically reactive component in the disposable reaction vessel during the direct flow-through, and in which the component to be determined is 2~3~00~
a) eluted and determined, or b) marked in the disposable reaction vessel via amplification reactions with common immunological markers, and then determined, or c) determined via a marked compound in reactive competition to the component to be determined.
The invention is described in detail with relation to figures 1 and 2 which show:
Fig. la a longitudinal section of a disposable reaction vessel 1 according to the invention containing two separating devices 2a and 2b, between which the substrate with the ' immunologically reactive component 3 is located, Fig. lb a top view of the disposable reaction vessel 1, and Fig. 2 a scheme of an online-system for antibody production control.
The dimensions given in figures la and lb are for example only.
The described disposable reaction vessel 1 and the described method, in which the disposable reaction vessel 1 is used, exhibit several considerable advantages compared to known reaction vessels resp. methods.
The described disposable reaction vessel 1 allows a rapid performance of the quantitative determination of components which can be determined by immunoreactions. The handling of the reaction vessel 1 is very simple and does not require the use of costly instruments such as HPLC apparatus. Besides excellent results are obtained with the aforesaid disposable reaction vessel 1 at extremely low consumption of substrate 3 with bound immunologically reactive component (named substrate bed in the following). R disposable reaction vessel 1 with a substrate bed volume 8, i.e. the volume between the upper and lower separating device 2a and 2b, of 50 ul is preferably used. The small substrate bed volume allows the use of small quantities of elution liquid with the consequence that the dilutio.- effect of the elution remains small, and thus the detection limit is not decreased in contrast to other methods in which larger elution volumes are necessary.
i`-'3` ~
Z131~0~7 Besides, the aforesaid reaction vessel 1 allows the quantitative determination at binding equilibrium (end-point determination) between the component to be determined and the immunologically reactive component. The binding equilibrium is reached within an incubation time of 1 minute already in the aforesaid disposable reaction vessel 1.
The rapid e~uilibration is rendered possible by the small substrate bed volume 8, its flow resistance, and the high loading of the substrate with the immunologically reactive component. The consequence of the controlled flow and the high loading of the substrate with the reactive component is that the component to be determined is bound quannatitatively to the reactive component within about 1 minute, i.e. before the flowing liqu;d leaves the substra,te bed volume. Therefore, a comparative analysis with standard references is spared.
Besides, a rapid analysis of samples is possible, because the disposable reaction vessel 1 allows immediate use, because the substrate 3 with the immunologically reactive component can be standardi~ed and ready-made, and the disposable reaction vessel 1 does not need to be calibrated before use.
The substrate 3 does not need to be stable against pressure, because the disposable reaction vessel 1 allows the rapid analysis of samples without application of HPLC. For the substrate 3, all common materials useful for affinity chromatography can be considered. Prefered substrate materials are polymeric sugars, plastics, plastic-modified substrates, metal oxides, or silicates.
Substrate materials in the form of frits are especially prefered.
For the determination of very large components such as viruses, which do not have access to the pores of such substrate materials, graft copolymers with side chains with active groups are prefered.
The immunologically reactive component loaded onto the substrate material 3 can bs bound to the substrate material covalently as well as adsorptively. The immunologically reactive component can be selected from the group containing haptens, antigens, antibodies, and immuno-affine proteins. Polyclonal as well as monoclonal antibodies can be used.
.. . . .
The vessel material 4 does not need to be stable against pressure, because the disposable reaction vessel 1 does not need to be subjected to high pressures in the present procedure. Therefore, besides pressure-resistant materials such as metals, other not pressure-resistant materials can be considered for the production of the disposable reaction vessel 1 such as glass, natural materials, and plastics. Prefered plastics are polyethylene~
polypropylene, and/or polystyrene, and prefered metals are aluminum and stainless steel.
For practi~cal purposes, the disposable reaction vessel 1 has a swelling 5 at one end to allow adaption of the reaction vessel into a holding and moving device for automatic transport. This is necessary for the use in automated sample processing devices.
~13~1007 ` - 5 -Besides, for practical purposes, the disposable reaction vessel 1 has a smaller-diameter joininy at the end opposite to the swelling, preferentially a joining ~or a socket-switch connection with another disposable reaction vessel~ This facilitates e.g.
joining in series of several disposable reaction vessels 1, whereby the outlet 6 of the first vessel is,connected with the inlet of the second vessel by a easy simple male-female connection.
Besides, the disposable reaction vessel 1 is closable by caps 7 and 9.
The present procedùre using said disposable reaction vessel 1 is superior due to, among others, a strongly simplified procedure compared to common procedures, and because it allows a rapid quantitative and qualitative determination. This is enabled partly because the determination of the component to be determined does not require a preceding calibration or regeneration of the used substrate in the disposable reaction vessel 1, and because no series reference measurements with standard solutions are ', necessary. The easy handling is a consequence of the possibility to apply samples and other solutions by usual laboratory equipment or by a pipetting automate. ~-Besides manual procedures, automated procedures are improved by application of the invented reaction vessel 1. In such procedures, the application of the sample and the further steps of the procedure ars performed by an automated sample processing device.
Such a sample processing device is used for example in the online production process control of antibodies produced in a fermenter.
In such procedures the samples are automatically applied in time intervalls of about 6 to 8 hours, and the antibody concentration in the fermentation medium can be continuously determined rapidly and easily over a long time period using the disposable reaction vessel 1. Q system for antibody production control using the invented disposable reaction vessel 1 is schematically described ~ ;
in fig. 2. Therein, the disposàble reaction vessel 1 is included in the automated sample preocessing device.
The present procedure can be performed rapidly without application of high pressures to the disposable reaction vessel 1 after sample application. But low suction or pressure forces can be used for quicker performance, e.g. by centrifugation or pumping.
If necessary, several disposable reaction vessels 1 can be connected in series or parallel in the present procedure. This allows e.g. the simultaneous determination of several parameters in one single sample resp. the simultaneous analysis of several samples.
.
2~3~007 The connection of several disposable reaction vessels in series is advantageous e.g. for the performance of allergy tests. Therein, a different antigen (allergen) is applied to each different column, then the columns are connected in series, and the sample containing the antibody of the class IgE which mediates the allergic reaction is applied to the first reaction vessel.
The sample flows successively through the different disposable reaction vessels 1 containing the different allergens. The IgE of the sample will be bound in the reaction vessel which contains the allergy-evoking antigen, and can be detected e.g. via fluorescence detection. In this way, one single sample can be tested simultaneously with many different potential allergens.
~ '''.;''' '` ~''''', ' ''' ; ' , ~ ' ;
Claims (28)
1. Disposable reaction vessel (1) for solid-phase immune analysis, open at the upper and lower end, with at least one immunologically reactive component loaded onto a substrate (3) with active or activated surface with such a loading density that the component to be determined is bound quantitatively (bonding equilibrium) to the reactive component, before the liquid flowing through the column leaves the substrate bed, wherein the substrate bed volume (8) is up to 600 ul, and wherein the flow-through velocity of the liquid is appropriately determined by the substrate bed.
2. Disposable reaction vessel according to claim 1, wherein the substrate (3) together with the immunologically reactive component is standardized and ready-made to allow immediate use of the disposable reaction vessel (1).
3. Disposable reaction vessel according to any one of claims 1 to 2, wherein the substrate material (3) is selected from the group containing polymeric sugars, plastics, plastic-modified organic or inorganic substrates, porous metals and alloys, metal oxides, glasses, silicates, or ceramic materials.
4. Disposable reaction vessel according to claim 3, wherein the substrate material is a solid frit or membrane, wherein the porosity of the substrate determines the flow-through velocity of the liquid according to claim 1.
5. Disposable reaction vessel according to one of claims 3 or 4, wherein the plastics is a modification of polyethylene, polypropylene, polyacrylate, polyamide, of any of their copolymers, or of cellulose, wherein the metal is aluminum, or where the metal oxide is aluminum oxide, titanium dioxide, or zirconium dioxide.
6. Disposable reaction vessel according to one of claims 1 or 2, wherein the substrate has a significant surface structure with a high concentration of active binding sites for the immunologically reactive component, and wherein the flow-through velocity of the liquid is determined by the relation between the particle size and the surface structure of the substrate according to claim 1.
7. Disposable reaction vessel according to claim 6, wherein the substrate is a graft copolymer (tentacle polymer), wherein the flow-through velocity of the liquid is determined by the relation between particle size and length of the side chains according to claim 1.
8. Disposable reaction vessel according to claim 1, wherein the immunologically reactive component is bound to the substrate (3) covalently or adsorptively.
9. Disposable reaction vessel according to claim 8, wherein the immunologically reactive component is selected from the group containing hapten, antigen, antibody, or immune-affine protein.
10. Disposable reaction vessel according to claim 9, wherein the antibody is polyclonal.
11. Disposable reaction vessel according to claim 9, wherein the antibody is monoclonal.
12. Disposable reaction vessel according to claim 4, wherein the substrate has a porosity of 0.2 um to 100 um.
13. Disposable reaction vessel according to claim 4, wherein the layer thickness of the substrate is 0.1 mm to 20 mm.
14. Disposable reaction vessel according to claim 7, wherein the graft copolymer is polystyrene with grafted polyethyleneglycol with a particle size of 10 to 15 um.
15. Disposable reaction vessel according to any of claims 1 to 14, wherein the vessel material is selected from the group containing plastics, glasses, metals, or natural materials.
16. Disposable reaction vessel according to claim 15, wherein the plastics is polyethylene, polypropylene and/or polystyrene.
17. Disposable reaction vessel according to any one of claims 1 to 16 with a swelling (5) at one end to allow the adaption of the reaction vessel into holding and moving devices for automatic transport.
18. Disposable reaction vessel according to any one of claims 1 to 17 with a joining (6) at the end opposite to the swelling (5) with a smaller diameter than the vessel diameter .
19. Disposable reaction vessel according to claim 18, wherein the reaction vessel includes male-female connecting elements at the upper and lower ends that permit connections between more than one vessel in a series.
20. Disposable reaction vessel according to any one of claims 1 to 19. wherein the substrate bed volume (8) is 50 ul.
21. Procedure for the determination of components which are determinable via an immunoreaction, comprising: applying a sample to be analyzed to a disposable reaction vessel according to any one of claims 1 to 20, wherein any of the component to be determined in the sample is bound to the complementary immunologically reactive component in the disposable reaction vessel, and determining the component to be determined a) via elution and determination;
b) via marking by amplifying reactions in the disposable reaction vessel with immunological markers and following determination; or c) via a compound competing with the compound to be determined.
b) via marking by amplifying reactions in the disposable reaction vessel with immunological markers and following determination; or c) via a compound competing with the compound to be determined.
22. Procedure according to claim 21, wherein the determination is made without preceding calibration or regeneration of the substrate and of the applied reactive component.
23. Procedure according to any one of claims 21 or 22, wherein the sample and the other solutions are applied with common laboratory equipment or with a pipetting automate.
24. Procedure according to any one of claims 21 or 22, wherein an automated sample processing device si used for sample application and further process steps.
25. Procedure according to claim 24, characterized by automated sample application in an on-line process control system.
26. Procedure according to any one of claims 21 to 25, wherein small suction or pressure forces are applied to the disposable reaction vessel after sample application for more rapid performance.
27. Procedure according to any one of claims 21 to 26, characterized by serial or parallel arrangement of several disposable reaction vessels.
28. Procedure according to claim 27, wherein the components to be determined are allergens.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DEP4208732.5 | 1992-03-18 | ||
| DE4208732A DE4208732C2 (en) | 1992-03-18 | 1992-03-18 | Disposable reaction vessel for solid phase immunoanalysis and method for measuring components that can be determined via immune reactions |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2131007A1 true CA2131007A1 (en) | 1993-09-30 |
Family
ID=6454401
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002131007A Abandoned CA2131007A1 (en) | 1992-03-18 | 1993-03-08 | One-way reaction vessel for the solid-phase immunological analysis of, and a method of measuring constituents which can be determined via immune reactions |
Country Status (17)
| Country | Link |
|---|---|
| EP (1) | EP0634015B1 (en) |
| JP (1) | JPH07504983A (en) |
| KR (1) | KR100270209B1 (en) |
| AT (1) | ATE158081T1 (en) |
| AU (1) | AU675781B2 (en) |
| BR (1) | BR9306086A (en) |
| CA (1) | CA2131007A1 (en) |
| CZ (1) | CZ217294A3 (en) |
| DE (2) | DE4208732C2 (en) |
| ES (1) | ES2106326T3 (en) |
| FI (1) | FI944320A0 (en) |
| HU (1) | HUT74558A (en) |
| NO (1) | NO943462L (en) |
| NZ (1) | NZ249491A (en) |
| RU (1) | RU2115122C1 (en) |
| SK (1) | SK109894A3 (en) |
| WO (1) | WO1993019368A1 (en) |
Families Citing this family (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1995019569A1 (en) * | 1994-01-13 | 1995-07-20 | Abion Beteiligungs- Und Verwaltungsgesellschaft Mbh | Reaction columns for simultaneous multiple measurement and method |
| DE19528847C2 (en) * | 1995-08-05 | 1997-12-11 | Biotest Ag | Affinity chromatographic method for the detection of antigens and / or antibodies |
| DE19543240A1 (en) * | 1995-11-20 | 1997-05-22 | Abion Ohg | Disposable applicator and kit |
| AU1597297A (en) * | 1996-01-30 | 1997-08-22 | Abion Beteiligungs- Und Verwaltungsgesellschaft Mbh | Carrier material loadable by a through flow for solid phase assays |
| GB9708961D0 (en) * | 1997-05-02 | 1997-06-25 | Ringel Karl Peter | Immunoassay |
| AU1315699A (en) * | 1997-11-12 | 1999-05-31 | Life Technologies, Inc. | Device and methods for purifying biological molecules using low volumes of chromatographic and adsorptive media |
| DE29803712U1 (en) * | 1998-03-04 | 1998-04-23 | Macherey, Nagel GmbH & Co. Handelsgesellschaft, 52355 Düren | Device for the treatment of biomolecules |
| US6177009B1 (en) | 1998-04-03 | 2001-01-23 | Macherey, Nagel Gmbh & Co. | Apparatus for treating biomolecules |
| DE10057195A1 (en) * | 2000-11-17 | 2002-05-23 | Robert Loesel | Reaction vessel, used for carrying out chemical reactions, comprises base formed by membrane having liquid permeability that can be controlled using pressures |
| DE10247430A1 (en) * | 2002-10-11 | 2004-04-29 | Fresenius Hemocare Gmbh | Method for determining the level of endotoxins in liquids |
| DE102007025311A1 (en) | 2007-05-30 | 2008-12-04 | Senova Gesellschaft für Biowissenschaft und Technik mbH | Quantitative heterogeneous fast binding assay with broad dynamic range and simple photometric detection |
| GB0802355D0 (en) * | 2008-02-08 | 2008-03-12 | Univ Greenwich | Modified SPE adsorbent cartridge |
| EP2264461A1 (en) | 2009-06-19 | 2010-12-22 | FZMB GmbH Forschungszentrum für Medizintechnik und Biotechnologie | Device for executing an affinity chromatographic sample test |
| DE11718963T1 (en) * | 2010-08-05 | 2015-10-01 | Vibod Gmbh | New columns for incubation and isolation of chemical and / or biological samples |
| EP2487490A1 (en) | 2011-02-11 | 2012-08-15 | FZMB GmbH Forschungszentrum für Medizintechnik und Biotechnologie | Heterogeneous binding assays with improved optical ability to be evaluated or porous fixed phase for same |
| JP6231989B2 (en) | 2011-11-10 | 2017-11-15 | バイオファイア・ダイアグノスティクス，リミテッド・ライアビリティ・カンパニー | Filling vial |
Family Cites Families (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE1907014A1 (en) * | 1969-02-12 | 1970-09-03 | Haller Dr Wolfgang | Process for separating blood components, in particular immunologically active globulins, from other components |
| US4039652A (en) * | 1973-10-11 | 1977-08-02 | Miles Laboratories, Inc. | Column method of immunoassay employing an immobilized binding partner |
| US4151254A (en) * | 1975-06-16 | 1979-04-24 | Union Carbide Corporation | Adsorption columns for use in radioimmunoassays |
| US4145406A (en) * | 1977-04-11 | 1979-03-20 | Miles Laboratories, Inc. | Specific binding - adsorbent assay method and test means |
| SE458643B (en) * | 1984-12-07 | 1989-04-17 | Pharmacia Ab | DEVICE WITH COLUMN ELEMENTS DIVIDED IN SAMPLE PARTY AND REFERENCE PATRIAR |
| JPH087217B2 (en) * | 1986-04-09 | 1996-01-29 | オリンパス光学工業株式会社 | Immunological measurement method applying affinity-chromatography |
| JPH07119769B2 (en) * | 1986-10-01 | 1995-12-20 | 株式会社日立製作所 | Automatic analyzer |
| DE3717211A1 (en) * | 1987-05-22 | 1988-12-01 | Diagen Inst Molekularbio | DEVICE AND METHOD FOR SEPARATING AND CLEANING MOLECULES |
| US4892710A (en) * | 1987-07-07 | 1990-01-09 | Bioprobe International, Inc. | Cartridge assembly with multi-purpose closure tubing |
| DD268161A1 (en) * | 1987-11-27 | 1989-05-24 | Staatliches Inst Fuer Immunpra | METHOD FOR THE PRODUCTION OF BACTERIAL ANTIGENES |
| GB8928501D0 (en) * | 1989-12-18 | 1990-02-21 | Unilever Plc | Reagents |
| US5183740A (en) * | 1990-02-23 | 1993-02-02 | The United States Of America As Represented By The Secretary Of The Navy | Flow immunosensor method and apparatus |
| DE4126436A1 (en) * | 1990-09-17 | 1992-03-19 | Abion Ohg | DISPOSABLE REACTION TUBE FOR SOLID-PHASE IMMUNAL ANALYTICS AND METHOD FOR MEASURING COMPONENTS DETERMINABLE BY IMMUNE REACTIONS |
-
1992
- 1992-03-18 DE DE4208732A patent/DE4208732C2/en not_active Expired - Fee Related
-
1993
- 1993-03-08 RU RU94041215A patent/RU2115122C1/en active
- 1993-03-08 AT AT93905315T patent/ATE158081T1/en active
- 1993-03-08 CZ CZ942172A patent/CZ217294A3/en unknown
- 1993-03-08 WO PCT/EP1993/000520 patent/WO1993019368A1/en not_active Application Discontinuation
- 1993-03-08 NZ NZ249491A patent/NZ249491A/en unknown
- 1993-03-08 DE DE59307345T patent/DE59307345D1/en not_active Expired - Lifetime
- 1993-03-08 BR BR9306086A patent/BR9306086A/en not_active Application Discontinuation
- 1993-03-08 KR KR1019940703222A patent/KR100270209B1/en not_active IP Right Cessation
- 1993-03-08 HU HU9402654A patent/HUT74558A/en unknown
- 1993-03-08 AU AU36316/93A patent/AU675781B2/en not_active Ceased
- 1993-03-08 SK SK1098-94A patent/SK109894A3/en unknown
- 1993-03-08 ES ES93905315T patent/ES2106326T3/en not_active Expired - Lifetime
- 1993-03-08 CA CA002131007A patent/CA2131007A1/en not_active Abandoned
- 1993-03-08 JP JP5516208A patent/JPH07504983A/en active Pending
- 1993-03-08 EP EP93905315A patent/EP0634015B1/en not_active Expired - Lifetime
-
1994
- 1994-09-16 NO NO943462A patent/NO943462L/en unknown
- 1994-09-16 FI FI944320A patent/FI944320A0/en not_active Application Discontinuation
Also Published As
| Publication number | Publication date |
|---|---|
| HU9402654D0 (en) | 1994-11-28 |
| HUT74558A (en) | 1997-01-28 |
| ES2106326T3 (en) | 1997-11-01 |
| NO943462D0 (en) | 1994-09-16 |
| BR9306086A (en) | 1997-11-18 |
| AU3631693A (en) | 1993-10-21 |
| WO1993019368A1 (en) | 1993-09-30 |
| CZ217294A3 (en) | 1995-02-15 |
| SK109894A3 (en) | 1995-05-10 |
| DE4208732C2 (en) | 1995-04-27 |
| FI944320A (en) | 1994-09-16 |
| DE4208732A1 (en) | 1993-09-30 |
| NZ249491A (en) | 1996-06-25 |
| AU675781B2 (en) | 1997-02-20 |
| FI944320A0 (en) | 1994-09-16 |
| RU94041215A (en) | 1996-10-10 |
| RU2115122C1 (en) | 1998-07-10 |
| ATE158081T1 (en) | 1997-09-15 |
| KR100270209B1 (en) | 2001-01-15 |
| EP0634015A1 (en) | 1995-01-18 |
| NO943462L (en) | 1994-09-16 |
| EP0634015B1 (en) | 1997-09-10 |
| KR950701076A (en) | 1995-02-20 |
| DE59307345D1 (en) | 1997-10-16 |
| JPH07504983A (en) | 1995-06-01 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| FZDE | Discontinued |