DE20215268U1 - Self-adhesive blotting membrane, useful in nucleic acid hybridization and immunoassay diagnostic tests, has reversible adhesive on the back face - Google Patents

Abstract

Self-adhesive blotting membrane (A) that is provided, on the back, with a reversible adhesive (I). Self-adhesive blotting membrane (A) that is provided, on the back, with a reversible adhesive (I). Blotting device that contains individual (A), reversibly glued, side by side, to a carrier.

Description

The present invention relates to a self-adhesive blot membrane which is provided with a reversible adhesive on the back.

Blotting, i.e. the transfer of electrophoretically separated biomolecules from an agarose or polyacrylamide matrix to a membrane, has developed into a standard procedure in molecular biology, especially in molecular diagnostics, since the mid-1970s.

Depending on the nature of the molecule to be transferred, a distinction is made between Southern blotting,

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Northern blotting and Western blotting. So-called Southern blotting was first described by E. Southern in 1975. In Southern blotting, DNA molecules that have previously been separated electrophoretically in polyacrylamide or agarose gels are usually transferred to a suitable membrane, such as a nitrocellulose membrane, using negative pressure. After transfer, the DNA is immobilized on the membrane. The covalent fixation of the DNA can be achieved by crosslinking the DNA using UV radiation or by baking at a temperature of around 80 ^° C. The DNA is then available for various applications, such as hybridization with a single-stranded DNA probe of a defined nucleotide sequence. The Southern blotting method is preferably used when it is to be determined whether or not a defined nucleotide sequence is present in the genomic DNA of an organism. Furthermore, the method can be used for restriction mapping of a gene or a gene segment as well as for elucidating the structure of a gene, e.g. by phage mapping.

In reference to the transfer of DNA molecules to membranes, the name Northern blotting has been established for the corresponding transfer of RNA. Northern blotting differs only slightly from Southern blotting due to the different properties of RNA and DNA. Northern blotting is mainly used in analyses that seek to qualitatively and/or quantitatively determine the expression of a specific gene in different tissues or cell lines at the transcription level. In Northern blotting, total RNA extracts or polyadenylated mRNA enriched using oligo(dT) affinity chromatography are generally used.

In addition to the classic Southern blotting and Northern blotting, so-called dot blotting and slot blotting methods are increasingly being used, in which the nucleic acid to be tested

Acid can be applied directly to the membranes in a gel without prior electrophoretic separation.

The detection of specific proteins or antibodies against such proteins in molecular diagnostics is usually carried out using the Western blotting method. In Western blotting, antigens and proteins such as glycoproteins or enzymes are transferred to a suitable membrane and immobilized, usually after electrophoretic separation in polyacrylamide gels. Based on specific binding properties, these proteins can be detected directly on the membrane using antibodies, lectins or enzyme substrates. Various methods are used for transfer, such as tank blotting or semi-dry blotting.

Furthermore, in protein analysis, methods analogous to the dot blot or slot blot methods described above are known, by means of which specific, purified proteins or polypeptides are applied to a corresponding membrane individually or as a defined mixture in an adjustable concentration. The application can be carried out, for example, using a stamp or contact-free using a nozzle.

Membranes that have been coated with one or more proteins/polypeptides as a sequence of closely spaced drops, so that the eye sees the shape of a sharply defined line, are known in specialist circles as so-called "line blots". The spectrum of molecules that can be used is broader than with classic Western blotting. For example, lipids can also be applied to a membrane using the technique described for subsequent detection using antibodies.

Western blot membranes or line blot membranes coated with defined antigens or a spectrum of antigens

have now found their way into routine diagnostics. For example, the detection of bacterial or viral proteins/polypeptides by incubating a suitably coated membrane using specific antibodies in a patient's blood or serum samples allows conclusions to be drawn about a possible acute or past bacterial or viral infection.

Protein-coated membranes suitable for pathogen diagnostics are commercially available, for example, from EUROIMMUN.

Since it is sufficient in most cases to detect only a small amount of the immobilized biomolecule using specific antibodies or specific DNA probes, blot membranes are regularly offered as membrane strips of only a few millimeters wide for cost reasons. This ensures that material consumption is kept low, since usually only a single sample (e.g. a patient's blood sample) is to be tested with a blot membrane.

The detection of a specific binding of an antibody or a nucleic acid probe to the biomolecule immobilized on the blot membrane is carried out in a multi-stage detection process depending on the type of the molecule.

All detection methods used in Southern blot, Northern blot or Western blot include steps in which the corresponding membranes (or membrane strips) are mixed with various washing or incubation solutions. For example, the membranes are covered with solutions containing BSA, casein or Tween to saturate non-specific binding sites. These solutions are then exchanged for various washing solutions.

Since all incubation steps are usually carried out by swirling the incubation vessel (usually a plastic tub), it has proven to be a major disadvantage in the state of the art that the blot membrane can float freely in the solution. This can lead to the side of the membrane coated with the corresponding biomolecule turning downwards during the filling, emptying or swirling of the incubation vessel and the coated side sticking to the bottom or the wall of the vessel. Under these conditions, optimal wetting of the coated side of the membrane with the incubation liquid is no longer guaranteed. This can lead to difficult to evaluate or even incorrect detection results.

Furthermore, when the incubation liquid is aspirated (either with a pipette or using the suction device of an incubator), the blot membrane can come into contact with the suction opening and block it. This means that the solution to be exchanged cannot be efficiently replaced with the new incubation solution, which can also lead to incorrect detection results. If the incubation vessels are emptied manually, there is also a risk that the membrane will be poured out of the incubation vessel together with the incubation solution to be exchanged, which could result in contamination of the membrane.

The disadvantages of conventional blot membranes known in the state of the art apply in particular to thin membrane strips, which, due to their low weight and the associated mobility in the corresponding incubation solutions, are subject to a particularly high risk of adhering to the vessel wall or being discarded with the incubation solution.

The object of the present invention was therefore to provide means by which the above-described disadvantages of the prior art in the detection of biomolecules immobilized on membranes are eliminated and with the help of which the reliability of the corresponding results is increased by avoiding artifacts.

According to the invention, this object is achieved by a self-adhesive blot membrane which is provided with a reversible adhesive on the back.

The reversible adhesive on the back of the blot membrane according to the invention allows the membrane to be fixed directly to the bottom of the incubation vessel, thereby preventing this membrane from turning over when the vessel is filled, emptied or swiveled during the incubation. The reversible adhesive can be applied over the entire extent of the corresponding blot membrane back or can be limited to a defined area of the blot membrane back (e.g. in the form of one or more dots or stripes).

In the context of the present invention, a reversible adhesive is understood to mean an adhesive that has reversibly adhesive properties, i.e. the blot membranes provided with the adhesive can be removed from a non-adhesive surface after contact with this surface (e.g. the bottom of a reaction vessel made of plastic or glass) without losing their adhesive properties. A reversible adhesive on the back of a blot membrane thus allows it to be repeatedly adhered to and detached from these non-adhesive surfaces. The reversible adhesive is also referred to below as a pressure-sensitive adhesive.

The blot membrane provided with the reversible adhesive does not lose its adhesive properties even if it is

I :

is wetted and/or covered with aqueous solutions, such as saline incubation solutions, for a longer period of time. This makes it possible, for example, to fix the corresponding blot membrane in different reaction vessels one after the other during the detection process and to stick it into a suitable protocol or laboratory book or a data sheet after the detection process has been completed.

The reversible adhesive applied to the back of the blot membrane according to the invention thus enables the membrane to remain in a designated location of the respective incubation vessel during the entire detection process, even when the incubation solutions and/or the incubation vessels are changed several times, and prevents the membrane from being poured out or turned over or sticking with its front side (i.e. the side on which the respective biomolecule was applied) to a wall of the reaction vessel or the like.

Any adhesive that allows the membrane provided with the adhesive to be repeatedly removed and reattached to a non-adhesive surface (e.g. made of glass or plastic) can be used as a reversible adhesive within the scope of the invention. The preferred working range of the reversible adhesive can be between 20 ^° C and 25 °C. However, it should also show a stable adhesive effect at higher temperatures, such as 50 ^° C-65 ^° C or more. Such reversible adhesives are known to those skilled in the art from the production of labels or so-called adhesive markers (e.g. Post-it*) and include, for example, the "Creativ Mount" adhesive from 3M. "Creativ Mount" does not lose its adhesive effect even at temperatures that are usually used in the context of nucleic acid hybridization (approx. 60°-80°). The adhesives used are usually referred to as "pressure sensitive adhesives". Preferably an adhesive is used that allows the membrane to be removed without damaging the membrane.

The reversible adhesive can either be applied directly to the back of the laminated membrane or, for example, in the form of a double-sided adhesive tape.

According to the invention, the blot membrane can have an antigen or a spectrum of antigens on the front side. In the context of the invention, an antigen is any biological molecule that can be specifically detected by means of its binding to an antibody. According to one embodiment of the invention, this antigen is a protein, a lipid, a nucleic acid (DNA/RNA), a carbohydrate, a complex molecule (such as a molecule that has a protein and/or a carbohydrate and/or a lipid portion, such as gangliosides, etc.) and/or an allergen. In the present case, an allergen is understood to be any substance that is capable of triggering hypersensitivity reactions (both of the immediate and delayed type) and a specific antibody-mediated immune reaction in the human or animal body. When detecting antibodies against nucleic acids in patient samples, such as when detecting systemic lupus erythematosus, an autoimmune disease characterized, among other things, by the presence of an antibody against double-stranded DNA, the antigen applied to the front side of the blot membrane according to the invention can be single-stranded or double-stranded DNA or RNA.

Since the blot membranes according to the invention have adhesive properties even when soaked when using the aqueous incubation solutions, it has proven to be irrelevant within the scope of the present invention whether the blot membrane is first introduced into the reaction vessel or whether the blot membrane is only attached to the bottom of the reaction vessel after the corresponding incubation liquid has been added.

According to a preferred embodiment of the invention, the front side of the blot membrane has one or more protein antigens. These proteins can be individual, isolated proteins, such as pathogen-specific proteins, such as affinity-purified herpes simplex virus 2 type-specific glycoprotein G2 (gG 2), Epstein-Barr virus capsid antigen, or defined mixtures of several proteins, such as protein-antigen extracts from Helicobacter pylori strains. The proteins can be purified from the corresponding pathogen strains (native antigens) or expressed in host cells and then purified (recombinant antigens).

Furthermore, the proteins can be human antigens, which can be used to detect antibodies in patient sera that indicate an autoimmune disease. For example, soluble liver antigen/liver-pancreas antigen (SLA/LP), native thyroglobulin, recombinant thyroid peroxidase or combinations thereof can be applied to the blot membranes according to the invention.

According to a preferred embodiment of the invention, the blot membrane has on its front side the protein spectrum of a human or animal tissue, a cell line or an infectious agent, from food or pollen (allergens), which was electrophoretically separated according to its molecular mass in a denaturing SDS polyacrylamide gel and transferred to the membrane by Western blotting. In this case, the "protein spectrum" is understood to mean the entire protein fraction of the respective cells or the respective cell compartment, which is immediately available for electrophoretic separation after their lysis. The respective sample can also contain other molecules such as lipids or nucleic acids, since these do not prove to be disruptive in the subsequent blot.

The antigens isolated from animal tissues can be, for example, proteins from the venom of bees and wasps, which can cause extremely severe allergic reactions in some people. The venoms can be obtained using methods that are well known in the art, e.g. by extracting venom sacs after freeze-drying wasps, or by electrical stimulation of living bees.

According to the invention, the tissues can be, for example, hepatic tissue, neuronal tissue, e.g. from the cerebellum, or tissue from the pancreas, kidney, eye, inner ear or skin. These tissues can be isolated from mammals, for example from primates or humans, using methods that are well known in the art. The protein fraction is usually isolated by lysing the cells (e.g. by repeated freezing and thawing or boiling) or by homogenizing the tissue and taking it up in a suitable buffer and, if necessary, purifying and/or concentrating the desired proteins/polypeptides.

The cell line can be any established cell line that produces one or more proteins specific to this cell line. In the context of the present invention, these are preferably human cell lines, for example cell lines that express one or more specific proteins for malignant and benign tumors. According to a preferred embodiment, the cell line is a human epithelioma cell line or the HeLa cell line (ATCC CCL 2 ; Gey et al. (1952) Cancer Res. 12, 264; Scheper et al. (2002) Autoimmunity reviews, Vol.1, Issues 1-2, p. 17 "Anti-Mi-2 Westernblot: A new test for serological detection of myositis autoantibodies"). According to a particularly preferred embodiment, the human epithelioma cell line is

The human epithelioma type 2 cell line HEp-2 (ATCC CCL 23, Moore et al. (1955) Cancer Res. 15, 598) is used. The protein fraction is obtained from these cell lines using methods well known in the art, whereby the lysis of the cells can be modified according to their origin.

The infectious agents can be any pathogenic agent, such as bacteria, viruses or eukaryotes. These infectious agents or antibodies against these pathogens can preferably be detected in body fluids such as serum, blood, plasma, joint aspirate, urine and cerebrospinal fluid. According to a preferred embodiment, the infectious agents within the scope of the present invention are, for example, human pathogenic Borrelia, such as Borrelia burgdorferi sensu lato (Borrelia burgdorferi sensu stricto, Borrelia garinii and Borrelia afzelii) and other human pathogenic infectious agents, such as Echinococcus granulosus, Epstein-Barr virus (EBV), Helicobacter pylori, hepatitis viruses, herpes simplex virus (HSV), human immunodeficiency virus (HIV), Treponema pallidum, Yersinia enterocolitica or cytomegalovirus (CMV). The relevance of protein antigens in the diagnosis of Borrelia is sufficiently described in the state of the art (see, for example, Ma et al., Serodiagnosis of Lyme Borreliosis by Western Immunoblot: Reactivity of various significant antibodies against Borrelia burgdorferi. J. Clin. Microbiol. Feb./1992: 370-376; Tilton and Ryan, The laboratory diagnosis of Lyme disease. J. Clin. Immunoassay. 16: 208-214 (1993); Hauser et al., Interpretation Criteria for Standardized Western Blots for Three European Species of Borrelia burgdorferi Sensu Lato. J. Clin. Microbiol. June/1997: 1433-1444; Wilske et al., MiQ 12/2000 Quality standards in microbiological-infectious diagnostics: Lyme borreliosis. Urban & Fischer Verlag. Munich, Jena. (2000)).

The proteins are extracted from these infectious agents using methods that are well known in the art. In the case of lysis of bacterial cells, this can be done, for example, by enzymatic degradation of the cell wall using lysozyme or lysostaphin (in Staphylococcus species) .

The detection of antibody binding to the corresponding antigens (e.g. proteins) can be carried out using methods that are adequately described in the state of the art (cf. F. Lottspeich, H. Zorbas "Bioanalytik", Spektrum Akademischer Verlag, Heidelberg, Berlin, 1998). For example, the antibody intended for detection can be directly coupled ("labeled") with a molecule that enables the detection of the antibody and (therefore also the bound antigen). Such molecules suitable for labeling, which are usually covalently bound to the antibody, have been described in large numbers in the field of molecular diagnostics and include, among others, fluorescent dyes such as fluorescein isothiocyanate or tetramethylrhodamine-5-isothiocyanate, luminescent dyes and radioactively labeled molecules. In addition, antibodies can also be used that are complexed with corresponding molecules suitable for labeling. (e.g. gold particles).

Enzymes by means of which certain chromogenic substrates or substrates whose conversion is accompanied by a change (e.g. oxidation) of a chromogenic molecule can be converted in a suitable manner are also considered as detectable molecules in the present case. For example, the antibody according to the invention can be coupled with a peroxidase, such as a horse radish peroxidase (HPR), or a phosphatase, such as an alkaline phosphatase. The detection of a reaction complex consisting of the labelled antibody and the bound antigen takes place depending on the molecule selected to label the antibody.

Methods for labelling antibodies are known to those skilled in the art and require no further explanation.

Alternatively, the reaction complex can also be detected in a two-step process using secondary antibodies. In this process, a primary, specific unlabeled antibody that binds to the antigen is incubated with a second, labeled antibody that is coupled to a corresponding detectable molecule and is directed against the primary antibody. The secondary antibody is then detected using the coupled molecule that serves as the label. In this indirect detection process, the molecules mentioned above can again be used for labeling. This two-step detection process is considerably more sensitive than the direct detection of the antibody, since several labeled secondary antibodies can bind to the primary antibody (signal amplification).

Such a two-stage detection method is used, among other things, in the serological examination of patients as part of pathogen diagnostics or in the diagnosis of pathological conditions that can be traced back to autoantibodies. The protein antigen or the spectrum of protein antigens on the membrane is used to examine the presence of correspondingly specific antibodies in the sample (e.g. serum, plasma, blood, urine, joint aspirate, cerebrospinal fluid, etc.). The antibody from the patient's serum that is bound to an antigen is then detected with a labeled anti-human antibody.

According to a further preferred embodiment of the present invention, the blot membrane has one or more nucleic acids for hybridization on the front side. This can be genomic DNA, plasmid DNA or DNA obtained by re-

striction enzymes. The DNA can be transferred from agarose gels or polyacrylamide gels to the blot membrane using conventional Southern blotting methods and subsequently immobilized by baking or UV crosslinking. It is also possible to apply the DNA to the membrane by dot blotting, for example by pipetting appropriately diluted DNA solutions directly onto the membrane or by applying it using a stamp or nozzle. The DNA immobilized on the blot membrane can then be hybridized with sequence-specific, labeled probes, e.g. with digoxigenin-11-dUTP 3'-labeled oligonucleotides. Corresponding hybrids can be visualized by methods that are sufficiently known in the state of the art (for example by detecting the digoxigenin-containing hybrids by means of chemiluminescence detection after addition of disodium 3-(4-methoxyspiro{l,2-dioxetane-3 , 2 ' - (5 ' -chloro) tricyclo [3 . 3 .1.1 ³ ' ⁷ ] decan} 4-yl)-phenylphosphate (CSPD; cf. "The DIG System User's Guide for Filtration Hybridization", Boehringer Mannheim, 1995).

Alternatively, the nucleic acid immobilized on the blot membrane can be RNA molecules. For example, total RNA preparations from various tissues and body fluids, such as from the brain, from tumors of the breast or uterus, etc., are available commercially (for example "Multiple Tissue Northern Blots", available from Sigma Aldrich). The detection of RNA molecules or parts thereof immobilized on the blot membrane is carried out, as described for DNA, using conventional methods, such as digoxigenin detection.

The usual detection methods for DNA or RNA hybrids include, similar to Western blotting, several washing or incubation steps in which the blot membrane is mixed with various washing solutions and/or incubation solutions.

According to the invention, it has therefore also proven advantageous in these detection methods to fix the blot membrane to the bottom of the respective reaction vessel during the incubation carried out by swirling/shaking.

Within the scope of the invention, blot membranes made of different materials can be used depending on the molecule to be detected. These membranes are preferably made of nitrocellulose, nylon, polyethersulfone or polyvinylidene difluoride or glass fiber material. Within the scope of the invention, membranes with a modified surface can also be used. Membranes are preferably used whose charge is adapted to the antigens, which ensures improved binding of the antigens to the membranes. By using positively or negatively charged membranes, the binding of correspondingly negatively or positively charged antigens is improved, while uncharged membranes can bind antigens independently of charge and thus also antigen mixtures with different charges. Furthermore, the membranes used can be hydrophobic or have different binding properties for different antibodies/antigens due to a variable pore size. In a further embodiment, the membranes consist of a material to which antigens bind covalently (e.g. the Immunodyne ABC membrane (PaII), a nylon membrane with covalent binding activity, or the UltraBind" membrane (PaII), a polyethersulfone membrane with functional aldehyde groups for the formation of Schiff bases with primary amino groups in proteins). The covalent binding of the antigens to the membranes is carried out by methods known in the art.

According to the invention, the membranes can be laminated to a suitable polymer carrier, such as a polymer film made of polyester, fine fabrics, or the like, to increase their stability and improve handling.

Lamination is described in detail in the prior art and is therefore well known to the person skilled in the art. According to a preferred embodiment, a blot membrane made of nitrocellulose is laminated onto a polymer film.

According to a further preferred embodiment of the present invention, the blot membranes described are reversibly glued to a suitable carrier. The carrier is intended to fix the blot strips and to provide them in an arrangement that is easy for the person skilled in the art to handle, preferably lying parallel next to one another. The membranes are preferably strip-shaped membranes of a small width. The membranes can thus be arranged next to one another like a matchbook and can be easily removed from this carrier immediately before use. Handling the strips is therefore made enormously easier, since they no longer have to be removed from a glass or plastic container with tweezers, which usually leads to an electrostatic charge on the membrane or even to its destruction (particularly when using fragile nitrocellulose membranes).

The number of membranes arranged on the carrier and preferably next to each other can in principle be chosen arbitrarily, but for reasons of better handling is usually between 5 and 40, preferably 8, 16, 24 or 30. The carrier onto which the blot membrane strips are glued consists, for example, of plastic or (coated) cardboard (such as coated cardboard) and has at least the spatial dimensions of the corresponding blot membranes. It should preferably be of sufficient mechanical stability in order to be able to offer appropriate protection to the usually fragile membrane strip.

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Figure 1: Preparation of the blot membrane strips according to the invention.

a) Apply the reversible adhesive to the back of the membrane

b) View of the front of the membrane after cutting the membrane into individual strips

c) View of the back of the membrane after cutting the membrane into individual strips

Figure 2: Fixation and subsequent incubation of the blot membrane strips according to the invention in an incubation tray designed for the simultaneous incubation of several strips.

a) Top view of the incubation tray with several incubation channels

b) Side view of the blot membrane strip fixed in the tray and an enlarged section of the contact point where the blot membrane strips are connected to the bottom of the incubation tray by means of the adhesive.

The invention is described below using examples.

Examples

1. Production of anti-HSV Western blot strips

The antigens of a complete extract of herpes simplex I viruses are separated by molecular mass using discontinuous polyacrylamide gel electrophoresis. The transfer is then carried out

of the antigens using a tank blotting process on a nitrocellulose membrane. Control band and affinity chromatographically purified glycoprotein G2 from HSV-2 are applied in lines to the membrane and a label with the batch number is stuck on. The nitrocellulose membrane is reinforced with a self-adhesive polyester carrier and coated on the back with a reversible spray adhesive (Creative Mount). The polymer-reinforced self-adhesive nitrocellulose membrane is then cut into 2 mm wide test strips by machine and the strips are glued into a transport packaging.

2. Using the blot membrane strips

The user takes a test strip from the packaging for each patient material to be tested (serum, plasma or other body fluids) and sticks the strip into the incubation channel using the reversible adhesive on the back. The incubation then takes place according to the test instructions (extract from the Anti-HSV Westernblot EUROIMMUN AG test instructions).

Blocking: Fill the incubation channels with 1.5 ml of Universal BufferPlus according to the number of serum samples to be tested. Remove the required number of blot strips from the packaging using tweezers and stick them directly into an incubation channel filled with buffer. The number on the strip must be legible. Incubate for 15 minutes at room temperature on a rocking shaker. Then drain the liquid completely from the channels.

Sample incubation: Add 1.5 ml of the diluted serum samples to each incubation well filled with a blot strip and incubate for 30 minutes on a rocker at room temperature.

Washing: Completely drain liquid from each well and wash for 3 x 5 minutes with 1.5 ml Universal BufferPlus each on a rocking shaker.

Conjugate incubation: Pipette 1.5 ml of enzyme conjugate solution (alkaline phosphatase-labeled anti-human IgG) into each incubation channel and incubate for 30 minutes at room temperature on a rocking shaker.

Washing: Drain all liquid from the gutters. Wash as above.

Substrate incubation: Pipette 1.5 ml of substrate solution into each incubation channel. Incubate for 10 minutes at room temperature on a rocking shaker.

Stopping: Completely drain liquid from each well and rinse each blot strip 3x1 minute with distilled or deionized water.

Evaluation: Glue the incubated blot strips into the evaluation protocol and carefully dab them with absorbent paper. Hold the evaluation template to the glued-on blot strips and align it so that the black bar above the number of the blot strip is in line with the positioning bar on the evaluation template. The batch number on the evaluation template must match the batch number on the blot strips. Clearly recognizable bands on the blot strips that match the markings on the evaluation template are entered in the evaluation protocol.

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Figure la: Western blot membrane (back) Figure Ib: Western blot membrane strip (front) Figure Ic: Western blot membrane strip (back)

Figure 2a: Western blot membrane strips in the incubation tray

(Top view) Figure 2b: Western blot membrane strips in the incubation tray

(Side view and enlarged detail)

Reference number :

(1): reversible adhesive

(2) : Western blot strips

(3) : Field for printing product specifications, e.g.

Batch number (4): Incubation tray (5): Western blot strips fixed in the tray with adhesive

Claims (16)

1. Self-adhesive blot membrane with a reversible adhesive on the back. 2. Blot membrane according to claim 1, characterized in that an antigen or a spectrum of antigens is applied to the front side. 3. Blot membrane according to claim 2, characterized in that the antigen is a protein, lipid, allergen, carbohydrate, sugar-containing structures and/or a nucleic acid. 4. Blot membrane according to claim 3, characterized in that one or more proteins are applied to the front side. 5. Blot membrane according to one of claims 1 to 4, characterized in that it has on the front side a protein spectrum of an animal, a human, a tissue, a cell line, an infectious agent, an allergen, such as food or pollen. 6. Blot membrane according to claim 5, characterized in that the tissue is from the liver, the cerebellum, the pancreas, the kidney, the eye, the inner ear or the skin. 7. Blot membrane according to claim 5, characterized in that the cell line is a human epithelioma cell line or the He-La cell line. 8. Blot membrane according to claim 7, characterized in that the human epithelioma cell line is the human epithelioma type 2 cell line HEp-2. 9. Blot membrane according to claim 5, characterized in that the infectious agent comprises human pathogenic Borrelia and other human pathogenic infectious agents. 10. Blot membrane according to claim 9, characterized in that the human pathogenic Borrelia are Borrelia burgdorferi sensu lato (Borrelia burgdorferi sensu stricto, Borrelia garinii and Borrelia afzelii). 11. Blot membrane according to claim 9, characterized in that the human pathogens are Echinococcus granulosus, Epstein-Barr virus (EBV), Helicobacter pylori, hepatitis viruses, herpes simplex virus (HSV), human immunodeficiency virus (HIV), Treponema pallidum, Yersinia enterocolitica or cytomegalovirus (CMV). 12. Blot membrane according to claim 1, characterized in that it has one or more nucleic acids for hybridization on the front side. 13. Blot membrane according to one of the preceding claims, characterized in that it consists of nitrocellulose, nylon, polyethersulfone or polyvinylidene fluoride. 14. Blot membrane according to one of the preceding claims, characterized in that it consists of nitrocellulose and is fleece-reinforced and/or laminated to a polymer carrier. 15. Blot membrane arrangement containing blot membranes according to claims 1 to 13, in which the individual blot membranes are reversibly glued next to one another on a suitable carrier. 16. Blot membrane arrangement according to claim 14, characterized in that the blot membranes are strips of small width.