EP2247626A1

EP2247626A1 - Novel matrix materials, methods for the production thereof, and use thereof in methods for isolating biomolecules

Info

Publication number: EP2247626A1
Application number: EP09712804A
Authority: EP
Inventors: Roland Fabis; Ralf Himmelreich
Original assignee: Qiagen GmbH
Current assignee: Qiagen GmbH
Priority date: 2008-02-22
Filing date: 2009-02-20
Publication date: 2010-11-10
Also published as: US20110105739A1; DE102008010693A1; WO2009103792A1

Abstract

The invention relates to functionalized matrix materials or matrices that have structures of general formula (I) T - (C=O)O - B - D, methods for the production thereof, and the use thereof in methods for purifying and isolating biomolecules.

Description

NΘUΘ Matrix materials, processes for their preparation and their use in processes for isolating biomolecules.

The present invention relates to functionalized matrix materials or matrices containing these materials which have structures of general formula I.

T - (C = O) O - B - D (I),

in the

T is a polymer or copolymer comprising units of

Acrylic acid and / or its derivatives

B is a covalent bond or an alkylene group of 1 to 6

Carbon atoms

D is a branched or unbranched C ₂ -C ₂ o-alkyl radical or C ₃ -C ₂ -

Cycloalkyrest whose carbon chain may optionally be interrupted by one up to 10 oxygen, sulfur or bivalent amino group (s) -NR ⁴ -, the same or different, and having one or more hydroxyl group (s), thiol group (s) or amino group (n), aldehyde group (s), carboxylic acid amide group (s), sulfonamide group (s) and / or halogen atom (s), or a group

-CH (XR ¹ ) - CH ₂ (YR ² ),

in the

X and Y are independently oxygen, sulfur or -NR ³ - and

^R1 and ^R2 are independently hydrogen or a branched or unbranched -C ₂₀ -alkyl radical or C ₃ -C ₂ -Cycloalkyrest, whose carbon chain, optionally interrupted by one to 10 oxygen Sulfuric or divalent amino group (s) -NR ⁴ -, which may be the same or different, and which is interrupted by one or more hydroxyl group (s), thiol group (s) or amino group (s), aldehyde group (s), carboxylic acid amide group (s) Sulfonamide group (s) and / or halogen atom (s) may be substituted,

R ^{3 is} hydrogen or a C 1 -C ₆ -alkoxy which is substituted by one or more

Hydroxyl group (s), thiol group (s) or amino group (s) and / or halogen atom (s) may be substituted,

R ^{4 is} hydrogen or a C 1 -C ₆ -alkyrestup with one or more

can mean

wherein, in the case that the carrier consists of a homopolymer of methacrylic acid, D may not have the meaning of -CH ₂ -CH ₂ OH.

Preference is given to structures of the general formula (I) in which

T is a polymer or copolymer comprising units of

Acrylic acid and / or methacrylic acid and / or their amides and / or their esters

B is a covalent bond or an alkylene group of 1 to 3

Carbon atoms

D is a branched or unbranched C ₂ -C ₀ -alkyl radical or C ₃ -C ₀ -

Cycloalkyrest, whose carbon chain may optionally be interrupted by one up to three oxygen, sulfur or bivalent amino group (s) -NR ⁴ -, the same or different, and the one or a plurality of hydroxyl group (s), thiol group (s) or amino group (s), aldehyde group (s), carboxylic acid amide group (s), sulfonamide group (s) and / or halogen atom (s), or a group -CH (XR ¹ ) - CH ₂ (YR ² ), in the

X and Y are independently oxygen, sulfur or -NR ³ -,

R ¹ and R ² are independently hydrogen or a branched or unbranched CRCI _O -alkyl radical or C ₄ -C ₀ -Cycloalkyrest, whose carbon chain optionally substituted by one or up to three oxygen, sulfur or divalent amino group (s) -NR ⁴ -, the same or different, and may be interrupted by one or more hydroxyl group (s), thiol group (s) or amino group (s), aldehyde group (s), carboxylic acid amide group (s), sulfonamide group (s) and / or halogen atom (s) ),

R ^{3 is} hydrogen or a C 1 -C ₆ -alkoxy which is substituted by one or more

R ^{4 is} hydrogen or a C 1 -C 6 -alkyl radical containing one or more

can mean

Particular preference is given to structures of the general formula (I) in which T is a polymer or copolymer comprising units of

Acrylic acid and / or methacrylic acid and / or their esters

B is a covalent bond or a methylene group

D is a branched or unbranched C ₂ -C ₈ -alkyl radical or C ₃ -C ₈ -

Cycloalkyrest, whose carbon chain may optionally be interrupted by one to two oxygen, sulfur or bivalent amino group (s) -NR ⁴ -, the same or different, and having one or more hydroxyl group (s), thiol group (s) or amino group (n), aldehyde group (s), carboxylic acid amide group (s), sulfonamide group (s) and / or halogen atom (s), or a group -CH (XR ¹ ) -CH ₂ (YR ² ) in which

X and Y are independently oxygen, sulfur or -NR ³ -,

R ¹ and R ² independently of one another denote hydrogen or a branched or unbranched C ₁ -C ₆ -alkyrest or C ₄ -C ₆ -cycloalkyne whose carbon chain may optionally be replaced by one or by up to two oxygen, sulfur or bivalent amino group (s). NR ⁴ -, the same or different, and may be interrupted by one or more hydroxyl group (s), thiol group (s) or amino group (s), aldehyde group (s), carboxamide group (s), sulfonamide group (s) and / or halogen atom (s) may be substituted,

R ^{3 is} hydrogen or a C 1 -C ₃ -alkyrest, containing one or more

R ^{4 is} hydrogen or a dC ₃ -alkyrest, which with one or more

can mean wherein in the case of the carrier consists of a homopolymer of methacrylic acid, D may not have the meaning of -CH ₂ -CH ₂ OH.

Very particular preference is given to structures of the general formula (I) in which

T is a crosslinked polymer consisting of units of

methacrylic

B is a covalent bond or a methylene group

D is -CH ₂ -CH (OH) -CH ₂ OH, -CH ₂ -C (CH ₂ OH) ₃ ,

-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -OH or a group -CH (XR ¹ ) -CH ₂ (YR ² ) in which

X and Y oxygen,

R ¹ and R ^{2 are} independently hydrogen, -CH (OH) -CH ₂ OH,

-CH ₂ -CH (OH) -CH ₂ OH, -CH ₂ -C (CH ₂ OH) ₃ , -CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -OH,

can mean.

Most preferred are structures of the general formula (I) in which

T is a polymer cross-linked with ethylene glycol dimethacrylate consisting of units of preferably one or more methacrylic acid ester (s)

B is a covalent bond or a methylene group D - CH ₂ -CH ₂ OH, -CH ₂ -CH (OH) -CH ₂ OH, -CH ₂ -C (CH ₂ OH) ₃ ,

-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -OH or a group -CH (XR ¹ ) -CH ₂ (YR ² ),

in the

X and Y oxygen,

R ¹ and R ^{2 are} independently hydrogen, -CH (OH) -CH ₂ OH,

can mean.

The present invention furthermore relates to processes for the preparation of the compounds according to the general formula (I) and their use in processes for purifying or isolating biomolecules.

In this context, biomolecules according to the present invention generally refer to molecules which, as products of evolutionary selection, perform specific tasks for an organism through ordered and selective interaction and form the basis of its vital functions [cf. Römpp, Lexicon Biochemistry and Molecular Biology, Georg Thieme Verlag, Stuttgart 1999].

In particular, for the purposes of the present invention, nucleic acids, such as e.g. Plasmid DNA, chromosomal DNA, total RNA, mRNA or RNA / DNA hybrids are considered biomolecules.

The use according to the invention relates to the purification or isolation of these biomolecules from aqueous systems which contain, for example, impurities such as fats or, inter alia, cell debris. In addition, the present invention relates to the isolation or purification of optionally artificial biomolecules from reaction mixtures resulting from laboratory or industrial synthetic reactions.

In particular, the invention relates to matrix materials or matrices which have the ability to reversibly immobilize nucleic acids under defined conditions. The invention also relates to processes for the preparation and use of such matrix materials for the production of magnetic matrices and their use for purifying or isolating the desired biomolecules - in particular of nucleic acid (s).

The tremendous scientific progress in the field of molecular biology - i.a. in the field of genetic engineering and molecular diagnostics - has created a high demand for fast, safe and automatable procedures for the isolation and purification of nucleic acids.

The biological samples, which i.a. used for DNA identification or analysis, derived from a variety of sources, e.g. Animal and plant cells, faeces, tissue and bone samples (biopsies) or blood derived. In addition, the samples can also be drawn from the soil, from food or from synthetically produced nucleic acid mixtures.

Many molecular biological methods, such as reverse transcription, cloning, restriction analysis, amplification and sequencing necessarily require that the nucleic acids used in the respective methods are essentially free of impurities which can adversely affect the corresponding reaction processes or analytical methods. Such contaminants generally include substances that catalyze or initiate the degradation or depolymerization of a nucleic acid, or substances that block the detection of the nucleic acid or mask the nucleic acid. Undesirable impurities also include macromolecular substances such as enzymes, proteins, polysaccharides or polynucleotides, as well as low molecular weight substances such as lipids, enzyme inhibitors or oligonucleotides. Other impurities are dyes (pigments), trace elements (metals) or organic solvents.

These requirements have led to the fact that in the prior art has already established a variety of purification and isolation methods over the years. Thus, e.g. DNA can be obtained by salting out in the presence of phenol and trichloromethane. On the other hand, DNA and RNA can be recovered by the use of chaotropic salts on mineral carriers (such as silica) or derivatized silica resins. In particular, the use of magnetic particles - the so-called 'magnetic beads' - has become established for use in automated processes.

First, Marko et al. [Analyte. Biochem. 121 (1982) 382] and Vogelstein et al. [Proc. Nat. Acad. Be. 76 (1979) 615] that if the DNA is exposed in nucleic acid-containing extracts to high concentrations of sodium iodide or sodium perchlorate, only the DNA binds to the mechanically finely divided glass scintillation vials and comminuted glass fiber or fiberglass plates, while RNA and proteins do not tie. The DNA thus bound can be eluted in the simplest case with water. Subsequently, numerous patents based on this type of DNA isolation were disclosed.

Thus, EP-A-389 063 relates to a method for isolating nucleic acids from a biological source. According to this method, the biological sources containing nucleic acids, such as blood, cells, plasma, etc., are digested in high concentrations in the presence of chaotropic salts. Subsequently, the nucleic acids are bound to a silica surface. The nucleic acids are then washed and eluted.

The process disclosed in US Pat. No. 5,155,018 describes the isolation of RNA from biological sources which contain not only RNA but also DNA and other ingredients. In this case, the biological sample is acidified and with a chaotropic agent such. As a guanidinium mixed. The sample Silicate particles are added. Under the given conditions, RNA binds to the silicate particles.

Subsequently, the RNA is separated from the particles in this process as well.

Colpan et al. describe in International Patent Application WO-A-95/01359 a method for purifying and separating nucleic acid mixtures by adsorption of the nucleic acid from a high ionic strength alcoholic solution. The adsorption solution contains in addition to alcohol in a concentration of 1 to 50 vol .-% salts in a concentration of 1 to 10 M, wherein the chaotropic salts, such as. As guanidinium thiocyanate, sodium perchlorate, or guanidinium hydrochloride are preferred.

The subject of WO-A-95/21849 is further a method for the separation of double and / or single-stranded nucleic acids from sources containing these nucleic acids. Also in this method, the nucleic acids are adsorbed on mineral supports under conditions that allow binding of the desired nucleic acid species while the unwanted nucleic acid species does not bind to that mineral support.

In order to bind mainly single-stranded nucleic acids to a mineral carrier and thus to separate from double-stranded nucleic acids, the treatment conditions for the samples containing both nucleic acid species are adjusted accordingly with an aqueous mixture of salts, in particular of chaotropic salts and alcohol. The unadsorbed double-stranded nucleic acid can then be further purified or isolated by known methods.

Further processes are described, for example, in European Patent Applications EP-A-512 767 and EP-A-515 484, in International Patent Applications WO-A-95/13368, WO-A-96/18731 and WO-A-97/10331 and US Pat in US Pat. Nos. 4,923,978 and 5,057,426.

European Patent Application EP-A-707 077 describes the binding as well as the subsequent selective release of nucleic acid from a lysate by means of a water-soluble weakly basic polymer that precipitates with the nucleic acid under specific conditions. This precipitate can be separated from the aqueous solution and the nucleic acid can be released using high salt concentrations as well as strong bases or by heating.

In addition, with regard to all isolation or purification procedures described above, it should be noted that blood is one of the largest sources of DNA analysis available, as blood samples are routinely drawn for a variety of reasons. Due to the viscous nature and the extremely protein-containing composition, the automation of the isolation of nucleic acids from this starting material repeatedly raised unexpected difficulties - a circumstance which has also been repeatedly proven in the prior art. In addition, it has proved difficult to handle large volumes of samples that have relatively small amounts of DNA in view of automation.

The present patent application is therefore based on the object at least partially to solve the problems described above and known from the prior art. Thus, the present invention, in the most general sense, addresses the need for materials and methods that enable a rapid and efficient method of isolating nucleic acids from a mixture of the desired nucleic acids with contaminants.

In addition, the patent application is based on the object to provide matrices or matrix materials available that are reusable after use.

This object is achieved in an embodiment according to the invention by functionalizing matrix materials of general formula I and by methods for isolating one or more nucleic acids, such as plasmid DNA, chromosomal DNA, total RNA, m-RNA, or RNA / DNA hybrids eg from biological samples Contaminants, such as proteins, fats, cell debris, etc., contained, dissolved. In a further embodiment of the invention, the present invention relates to a modified matrix of the general formula I and to processes for preparing the matrix materials of the general formula I or matrices which comprise these matrix materials.

According to a further aspect, the invention relates to a method for isolating biomolecules - in particular nucleic acids - using the above-mentioned matrix, which comprises the following steps:

(a) providing the matrix according to the invention;

(b) combining the matrix with a mixture containing the biomolecule to be isolated or preferably containing the nucleic acid to be isolated in addition to at least one contaminant;

(c) incubating the matrix and the mixture under conditions under which the biomolecule or preferably the desired nucleic acid is immobilized on the matrix;

(d) separating the matrix with the biomolecule or immobilized nucleic acid (s) from the mixture;

(e) combining the matrix with the biomolecule or with the immobilized nucleic acid (s) with an elution solution, the desired biomolecule (s) or nucleic acid (s) being desorbed from the matrix.

The synthesis of structures corresponding to general formula I is well-known from the prior art, starting from the corresponding epoxides, since, for example, epoxy resins containing epoxide and oxirane partial structures, respectively, are thus a suitable starting material for the preparation of Compounds that fall under the general formula I - embodying products that are used on a large industrial scale (eg in the paint industry). The corresponding materials of the general formula I according to the invention can be readily made available in a subsequent reaction step by opening the epoxide ring (inter alia, for example by way of hydrolysis or alcoholysis).

In general, the structures of general formula I of the invention can be prepared from precursor molecules having suitable precursor groups in numerous ways known in the art.

Thus, e.g. suitable oxiranes can be prepared by the epoxidation (Prileschajew reaction) of alkenes or of hydroperoxides during the autoxidation of olefins. In addition, the desired in the intermediate epoxide partial structures starting from suitable precursor groups by dehydrohalogenation of halohydrins or by the reaction of carbonyl compounds with sulfur yelide nucleophiles (Corey) produce. For further explanation, it should be noted that such structures are also accessible by the epoxidations of allylic alcohols (Sharpless-Katsuki epoxidation) or of olefins with sodium hypochlorite under manganese (III) catalysis (Jacobsen-Katsuki epoxidation).

Furthermore, the reaction of α-haloesters with carbonyl compounds in the presence of sodium ethoxide gives access to the corresponding structurally related 2- (ethoxycarbonyl) oxiranes (Darzens reaction).

For the purposes of the present invention, however, it is preferred to incorporate epoxi-functionalized monomers into the polymer forming the carrier, so that even the polymerization yields an intermediate product which has the desired oxirane partial structure and which in the subsequent step involves ring opening of the epoxide ring - e.g. by acid hydrolysis or alcoholysis - can be converted into the desired end product of general formula I.

Preferably, a based on acrylic acid derivatives terpolymer in which the epoxy partial structures by copolymerization of a Oxirane partial structure containing monomer and optionally another monomer which serves as a cross-linker, a beneficial intermediate.

Particularly preferably, glycidyl methacrylate may be added as the epoxy-functionalized monomer in the polymerization of methacrylates, preferably in the presence of glycol dimethacrylate which serves as cross-linker. The resulting polymer thus has the required for the preparation of the compounds of the general formula I oxirane partial structures (X = O and R ¹ forms with R ² and Y is a single bond), by the subsequent hydrolysis / Akoholyse - or on the Ways of other known from the prior art reactions - can be easily converted into the target compounds of general formula I.

Such a functionalized polymer based on methacrylic acid derivatives - in particular methacrylic acid esters and methacrylic acid glycidyl esters - represents according to the invention a particularly preferred intermediate product.

Matrix materials of the general formula I can furthermore be prepared by simple transesterification reactions - preferably of acrylic or methacrylic acid esters which have a short-chain alkanol partial structure. Examples of such starting materials are the esters of acrylic or methacrylic acid with dC ₃ - called alcohols.

The geometric shape in which the functionalized polymer (s) or the matrix materials according to the invention are / is present plays a largely irrelevant role. For example, the surfaces may be in the form of particles, membranes, fleeces or fabrics.

The matrix can also be present as a so-called composite in which the matrix material is present, for example, in combination with an inorganic polymer such as silica (SiO ₂ ) or on a metallic surface such as gold or iron, or in any other Form combined with her. Accordingly, the term "matrix" in the context of the invention is to be understood as a matrix material if the latter singular is present, but matrix also denotes the combination of the matrix materials according to the invention with other components, such as, for example, the above-mentioned composites.

In general, it is possible for the preparation of the matrix to apply the functionalized polymers (matrix materials) to any hydrophilic or hydrophobic, possibly precoated, surface, resulting in a wide range of applications (inter alia in multi-well systems such as microtiter plates or microfluidic systems).

For the purposes of the invention, the support (T) of the matrix material or the matrix is formed by a polymer, the term polymer being understood in its broadest definition. According to the invention, polymers in the context of the present invention embody polymers, polycondensates or polyadducts. According to the present invention, the carrier may be composed of homopolymers, copolymers, terpolymers, etc. or of mixtures of various polymeric constituents. The optionally different polymers can - if desired - be crosslinked with each other with a so-called. Crosslinker. The term "carrier" for the purposes of the invention denotes the carrier of the functional group - (C = O) O-B-D.

For the preparation of the matrix material, suitable monomers which have at least one partial structure which are suitable for polymerization (or polycondensation or for the formation of polymeric adducts) and which have a second partial structure which has a precursor function for forming the partial structure - (C = O) OBD has. With suitable groups (B and D) in the starting material it may be necessary to dispense with the detour via a precursor. This may be the case, for example, if this group (s) does not adversely affect / influence the polymerization or is not impaired under the reaction conditions of the polymerization in its later function.

According to the invention, the abovementioned substituents, unless stated otherwise, have the following meaning in the context of the present invention: C 1 -C ₆ -alkyl, also generally referred to as "alkyl", is generally a monovalent, branched or unbranched hydrocarbon radical having 1 to 6 carbon atoms, optionally with one or more halogen atoms - preferably fluorine or - may be substituted by one or more hydroxy groups (n), which may be identical or different from one another, for example the following unsubstituted hydrocarbon radicals:

Methyl, ethyl, propyl, 1-methylethyl (/ so-propyl), butyl, 1-methylpropyl, 2-methylpropyl, 1, 1-dimethylethyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, 1, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, hexyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 1, 1-dimethylbutyl, 1, 2-dimethylbutyl, 1,3-dimethylbutyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, 3,3-dimethylbutyl, 1-ethylbutyl, 2-ethylbutyl, 1, 1, 2-thymethylpropyl, 1, 2,2-thymethylpropyl, 1 - Ethyl 1-methylpropyl and 1-ethyl-2-methyl-propyl.

The same applies to other - for example, smaller or larger - alkyl radicals, such as C _r C ₃ alkyl or C ₄ -C ₆ alkyl.

Higher alkyl radical is a monovalent branched or unbranched C ₇ - C ₂ o-alky radical which may optionally be substituted by one or more halogen atom (s), preferably fluorine, which may be identical or different. Examples which may be mentioned are the following preferred unsubstituted hydrocarbon radicals: branched or unbranched heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tetradecyl, hexadecyl, dodecadecyl and eicosyl.

Alkylene generally represents an unbranched, divalent hydrocarbon radical of 1 to 6 carbon atoms or a branched, divalent hydrocarbon radical of 3 to 6 carbon atoms, such as e.g. Methylene, ethylene, propylene, 2-methylpropylene, pentylene and the like.

Cycloalkyl is, unless defined otherwise, a saturated monovalent cyclic hydrocarbon radical having from three to eight carbon atoms and optionally with one or more halogen atoms - preferably fluorine - or may be substituted with one or more hydroxy group (s), which may be the same or different. Examples include: cyclopropyl, cyclohexyl, cycloheptyl or cyclooctyl. In this case, the carbon chain can be interrupted by one or more heteroatoms, such as, for example, nitrogen, oxygen or sulfur. Examples which may be mentioned are 1, 4-dioxixane (dioxane), 2,5-dihydrofuran, tetrahydrofuran, γ-pyran, 2H-1,3-dioxole, tetrahydropyrrole, 2,5-dihydropyrrole, piperidine, piperazine, tetrahydrothiophene, morpholine, 1 , 2-oxathiolane, 1, 3-thiazole, 1 H-4,1, 2-thiadiazine, 1, 2-oxathiepan, γ-thiopyran, 1, 4-thiazixin (1, 4-thiazine).

As preferred examples of oxygen-containing, heterocyclic and hydroxy substituted cyclic systems may be mentioned i.a. for example, the carbohydrates (pentoses as well as hexoses), such as: arabinoses, xyloses or riboses or glucoses, mannoses, galactoses, fructoses or sorboses.

Halogen is - unless otherwise stated - fluorine, chlorine, bromine or iodine, but preferably fluorine and chlorine.

In the context of the present invention, support material means the polymeric backbone which is built up, if appropriate, by the polymerization of the starting monomers with a cross-linker and, if appropriate, other auxiliaries. By auxiliaries are meant according to the invention primarily substances that influence, for example, on the chemical or physical properties of the polymer. These include, for example, pore formers, which influence the porosity of the polymer, or substances which impart magnetic properties to the polymer. The carrier material can thus - as desired - be connected to a material having a permanent or temporary magnetic moment.

The present invention further relates to a process for the purification or isolation of nucleic acids comprising: providing the modified matrix according to the invention which is to be used in the process. The subsequent immobilization step consists in combining the matrix with the mixture containing the nucleic acid to be isolated in addition to at least one contaminant, under conditions in which the desired Nucleic acid is adsorbed to the matrix, separating the matrix with the immobilized nucleic acid from the mixture and desorbing the nucleic acid, wherein the desired nucleic acid (s) is (are) desorbed from the matrix.

The exact reaction conditions - which are necessary to ensure adsorption and desorption of the nucleic acids on or from the matrix - depend on various factors. These include essentially the nature of the matrix, the nature of the desired nucleic acid (DNA or RNA, molecular weight and nucleotide sequence composition, pK _b and pK _{a of} the basic or acidic partial structures, the density of these structures on the surface and last but not least the ability of the matrix to bind to the nucleic acid (s)).

Furthermore, it is not excluded that impurities in the mixture upon adsorption of the nucleic acid (s) adversely interfere with the binding step.

The mixtures used here which contain the desired nucleic acids can, on the one hand, be prepared from synthetically prepared reaction mixtures, as described, for example, in US Pat. incurred in the PCR - or - as already mentioned - come from biological samples.

As nucleic acid (s) containing biological samples are - for the purposes of the present invention - cell-free sample material, plasma, body fluids, such as blood, serum, cells, leukocyte fractions, Crusta phlogistica, sputum, urine, semen, faeces, smears, punctates, tissue samples each Type - such. As biopsies -, tissues and organs, food samples containing free or bound nucleic acids or nucleic acid-containing cells, environmental samples containing free or bound nucleic acids or nucleic acid-containing cells - such. B. Organisms (single or multi-celled insects, etc.), plants and parts of plants, bacteria, viruses, yeasts and other fungi, other eukaryotes and prokaryotes, etc. understood. When using such samples, however, it is first necessary to destroy the cells in order to make the nucleic acids accessible for the adsorption step.

If the desired nucleic acid is RNA, the adsorption conditions must be adjusted to promote adsorption of the desired RNA. If, in addition to the RNA, DNA is also present in the mixture, the adsorption conditions can be adjusted such that the binding of a species, in this case in particular RNA, to the matrix is preferably promoted. However, the specific adsorption conditions required to allow the adsorption and ultimately the desorption of RNA depend on the properties of the matrix and must be determined for each different type of matrix.

The matrix used in the invention is preferably in a form which can be separated from the mixture containing the desired nucleic acid, among other substances / impurities, by the action of an external force after the mixture has been mixed with the matrix. It will be understood by those skilled in the art that the type of external force that is suitable for separating the matrix from the mixture depends on the shape and physical properties of the matrix. For example, in the simplest case, the separation may be under the action of gravity, for example when the matrix is in the form of the solid phase of a chromatographic separation system.

On the other hand, the matrix material can be added to the mixture of the nucleic acid and impurity (s), if appropriate batchwise, and then separated again by decanting or filtering.

The external force which can be used in the isolation method of the invention, inter alia, may be a liquid under high pressure, the matrix forming the stationary phase of high pressure liquid chromatography. Other forms of external forces suitable for use in the method proposed by the invention include vacuum filtration, centrifugation or, preferably, magnetic force.

When using magnetic force, matrix materials or matrices which have a permanently or temporarily magnetic material are preferred. In the present invention, particles are preferably ferromagnetic or ferrrimagnetische used, preferably selected from the group consisting of: γ-Fe ₂ O ₃ (maghemite), Cr ₂ O _3, as well as ferrites, in particular of the type (M ²⁺ O) Fe ₂ O ₃ , where M ^{2+ is} a divalent transition metal cation and preferably Fe ₃ O ₄ (magnetite). However, other ferromagnetic or ferrimagnetic particles may also be used. These particles have an average particle diameter of less than 5 .mu.m, preferably less than 1 .mu.m and more preferably in an interval between 0.05 and 0.8 .mu.m and most preferably in a range of 0.1 and 0.4 .mu.m.

Examples of suitable commercially available ferro- or ferrimagnetic particles are magnetic particles on the basis of γ-Fe ₂ O ₃ - as Bayoxide ^® E AB 21, on the basis of ferrimagnetic magnetite as type Bayoxide ^® E 8706, E 8707, E 8710 and E 8713H (available from Lanxess AG, Leverkusen, DE) and as Magnetic Pigment 340 and as Magnetic Pigment 345 (available from BASF AG, Ludwigshafen am Rhein, DE).

In addition, superparamagnetic materials can also be used. As superparamagnetic materials are Fe, Fe ₃ O ₄ , Fe ₂ O ₃ , superparamagnetic ferrites Co, Ni, and binary and or ternary compounds (alloys) in question.

By way of example, iron oxide crystals having a diameter of 300 angstroms and smaller may be mentioned here.

The following examples are intended to illustrate the invention without limiting it. 1 . Preparation of Modified Beads Having a Polymethacrylate Surface Having Functional Groups of General Formula I.

[D = CH (XR ¹ ) - CH ₂ (YR ² ) with X and Y = O; R ¹ and R ² = H (1.1) or

-CH ₂ -C (CH ₂ OH) ₃ CJ.- ?;]:

1 .1 Hydrolysis of the epoxide

5 g of epoxy-functionalized magnetic polymer prepared by polymerization of methyl methacrylate using ethylene glycol dimethacrylate as a cross-linker and glycidyl methacrylate as the carrier of the epoxide functionality are dissolved in a 250 ml flask in 100 ml of 100 mM aqueous hydrochloric acid. The reaction mixture is degassed twice under a water-jet vacuum (rotary evaporator) and stirred for a period of about 12 hours at a temperature of 60 ° C. The reaction mixture is poured into a suction filter and the modified magnetic polymer particles are washed twice each with deionized water. washed twice with 10 mM hydrochloric acid, twice with 100 mM aqueous sodium phosphate solution (pH = 6) and last twice with deionized water and then resuspended in 100 ml of deionized water.

1 .2 alcoholysis of the epoxide

5 g of epoxy-functionalized magnetic polymers prepared by polymerization of methyl methacrylate using ethylene glycol dimethacrylate as a "cross-linker" and glycidyl methacrylate as the carrier of the epoxide functionality are suspended in a 250 ml flask in 100 ml of a 100 mM aqueous hydrochloric acid solution The reaction mixture is degassed twice under a water-jet vacuum (rotary evaporator) and then stirred for a period of about 12 hours at a temperature of 70 ° C. The reaction mixture is poured into a suction filter and the modified magnetic polymer particles are washed four times with deionized water and then resuspended in 100 ml of deionized water. 2. Preparation of Modified Beads Having a Polymethacrylate Surface Having Functional Groups of General Formula I.

[D = -CH ₂ -CH (OH) -CH ₂ (OH) (2.1) or -CH ₂ -C (CH ₂ OH) ₃ (2.2)]:

2.1 transesterification of methyl methacrylate with glycerol

15 ml of a suspension of a magnetic polymer consisting of methyl methacrylate and ethylene glycol dimethacrylate or divinylbenzene, in ethanol, are placed in a glass filter with porosity 4 and washed four times with anhydrous diglyme or toluene. The residue is taken up in a mixture of 5 g of glycerol in 45 ml of diglyme or toluene and the mixture is transferred to a 100 ml three-necked flask. The flask is then fitted with a reflux condenser and the suspension is degassed twice. With stirring (100 rpm), the reaction mixture is subjected to transesterification at a reaction temperature of 120 ° C. over a period of 16 hours. Thereafter, the magnetic polymers are washed four times with deionized water and three times with ethanol and stored under ethanol.

2.2 Transesterification of methyl methacrylate with pentaerythritol

15 ml of a suspension of a magnetic polymer consisting of methyl methacrylate and ethylene glycol dimethacrylate or divinylbenzene, in ethanol, are placed in a glass filter with porosity 4 and washed four times with anhydrous diglyme or toluene. The residue is taken up in a mixture of 5 g of pentaerythritol in 45 ml of diglyme or toluene and the mixture is transferred to a 100 ml three-necked flask. The flask is then fitted with a reflux condenser and the suspension is degassed twice. With stirring (100 rpm), the reaction mixture is subjected to transesterification at a reaction temperature of 120 ° C. over a period of 16 hours. Thereafter, the magnetic polymers are washed four times with deionized water and three times with ethanol and stored under ethanol. 3. Purification of DNA with Diol-Modified (Hydrolysis) Magnetic Polymers

Using two of the four buffer systems listed below, two DNA samples were purified. The average yield was determined. Diol-modified particles (see 1 .1!) Were used as magnetic particles.

a) 190 μl buffer RLT (QIAGEN) [guanidinium thiocyanate-containing buffer (pH 6.7-7.2)] and 225 μl ethanol b) 230 μl buffer P1 / P2 / P3 (QIAGEN) and 175 μl ethanol [buffer P1:

Chelating agent-containing buffer in the range of pH 7.8 to 8.2; Buffer P2: a detergent-containing alkaline lysis buffer (pH 13); Buffer P3: acetate-containing buffer (pH 5.4 to 5.6).

c) 190 μl buffer RLT (QIAGEN) and 100 μl diglyme d) 190 μl buffer RLT (QIAGEN) and 100 μl triglyme e) 190 μl buffer RLT (QIAGEN) and 100 μl 1, 4-dioxane

After the addition of 20 μl bead suspension (corresponding to 49 mg beads per ml), the mixture was incubated at 1100 rpm over a period of 2 min by means of a thermal vibrator. The reaction mixture was separated using magnetic force and the supernatant discarded. Thereafter, 400 μl of wash buffer PE (QIAGEN) [TRIS-containing buffer (pH 7.9-8.1)] was added and the reaction mixture was incubated for 1 min at 1100 rpm using a thermo-agitator. The reaction mixture was then separated using magnetic force and the supernatant removed. This washing step was repeated. After removing the supernatant, the reaction vessels were dried over a period of 10 minutes to remove residual ethanol. Thereafter, 300 μl of RNase-free water was added. The resulting suspension was mixed by means of a thermal vibrator over a period of one minute. The suspension was separated again using magnetic force and the supernatant was discarded. Subsequently, 100 μl of the aqueous solution of a 100 mM TRIS buffer (pH = 9.5) was added. Thereafter, the suspension was mixed over a period of 1 minute at 1100 rpm by means of a thermal shaker. Elution was performed using magnetic force and a magnetic separator. Finally, the elution step was repeated and the DNA content was determined by an OD determination and by gel electrophoresis, the following results being obtained:

Buffer system RSD 001 [μg DNA] RSD 002 [μg DNA]

A 0.98 0.82

B 1, 36 1, 54

C 2.44 3.62

D 4.02 8.71

E 1, 45 1, 50

The present invention thus relates to matrix materials according to the general formula I.

T - (C = O) O - B - D (I) in which

T is a polymer or copolymer comprising units of acrylic acid and / or derivatives thereof

B is a covalent bond or an alkylene group having 1 to 6 carbon atoms

D is a branched or unbranched C ₂ -C _2O -AI kyrest or C ₃ -C ₂ -

Cycloalkyrest, whose carbon chain optionally by a up to 10 oxygen, sulfur or bivalent amino group (s) -NR ⁴ -, the same or differently, and may be substituted with one or more hydroxyl group (s), thiol group (s) or amino group (s), aldehyde group (s), carboxylic acid amide group (s), sulfonamide group (s) and / or halogen atom (s) can or a group

-CH (XR ¹ ) - CH ₂ (YR ² ),

in the

X and Y independently of one another oxygen, sulfur or -NR ³ -

and

R ¹ and R ² are independently hydrogen or a branched or unbranched Ci -C _2O -AI kyrest or C ₃ - C ₂ -Cycloalkyrest whose carbon chain optionally interrupted by one to 10 oxygen, sulfur or divalent amino group (s) -NR ⁴ -, the same or different, and may be interrupted by one or more hydroxyl group (s), thiol group (s) or amino group (s), aldehyde group (s), carboxylic acid amide group (s), sulfonamide group (s) and / or halogen atom (s) ),

R ^{3 is} hydrogen or a C 1 -C ₆ -alkoxy which may be substituted by one or more hydroxyl group (s), thiol group (s) or amino group (s) and / or halogen atom (s),

R ^{4 is} hydrogen or a C 1 -C ₆ -alkoxy radical which may be substituted by one or more hydroxyl group (s), thiol group (s) or amino group (s) and / or halogen atom (s), can mean

wherein in the case of the carrier consists of a homopolymer of methacrylic acid, D may not have the meaning of -CH ₂ -CH ₂ OH.

The present invention relates in particular to matrix materials according to the general formula I.

T - (C = O) O - B - D (I) in which

T is a polymer or copolymer comprising units of

B is a covalent bond or an alkylene group having 1 to 3 carbon atoms

D is a branched or unbranched C ₂ -C ₀ -alkyl radical or C ₃ -C ₀ -

Cycloalkyrest whose carbon chain may optionally be interrupted by one up to three oxygen, sulfur or bivalent amino group (s) -NR ⁴ -, the same or different, and having one or more hydroxyl group (s), thiol group (s) or amino group (n), aldehyde group (s), carboxylic acid amide group (s), sulfonamide group (s) and / or halogen atom (s), or a group

-CH (XR ¹ ) - CH ₂ (YR ² ),

in the

X and Y are independently oxygen, sulfur or -NR ³ -, R ¹ and R ² independently of one another denote hydrogen or a branched or unbranched C 1 -C 10 -alkoxy radical or C ₄ -C 10 -cycloalkyl radical, the carbon chain of which is optionally substituted by one or up to three oxygen, sulfur or bivalent amino group (s) -NR ⁴ -, same or different, and may be interrupted by one or more hydroxyl group (s), thiol group (s) or amino group (s), aldehyde group (s), carboxylic acid amide group (s), sulfonamide group (s) and / or halogen atom ( en) can be substituted,

R ^{4 is} hydrogen or a C 1 -C ₆ -alkoxy radical which may be substituted by one or more hydroxyl group (s), thiol group (s) or amino group (s) and / or halogen atom (s),

can mean

The present invention preferably relates to matrix materials according to the general formula I.

T - (C = O) O - B - D (I) in which T is a polymer or copolymer comprising units of acrylic acid and / or methacrylic acid and / or their esters

B is a covalent bond or a methylene group

D is a branched or unbranched C ₂ -C ₈ -alkyl radical or C ₃ -C ₈ -

Cycloalkyrest, whose carbon chain may optionally be interrupted by one to two oxygen, sulfur or bivalent amino group (s) -NR ⁴ -, the same or different, and having one or more hydroxyl group (s), thiol group (s) or amino group (n), aldehyde group (s), carboxylic acid amide group (s), sulfonamide group (s) and / or halogen atom (s), or a group

-CH (XR ¹ ) - CH ₂ (YR ² ), in the

X and Y are independently oxygen, sulfur or -NR ³ -,

R ¹ and R ² independently of one another denote hydrogen or a branched or unbranched C ₁ -C ₆ -alkyrest or C ₄ -C ₆ -cycloalkyne whose carbon chain may optionally have one or up to two oxygen, sulfur or bivalent amino group (s). NR ⁴ -, the same or different, and may be interrupted by one or more hydroxyl group (s), thiol group (s) or amino group (s), aldehyde group (s), carboxamide group (s), sulfonamide group (s) and / or halogen atom (s) may be substituted,

R ^{3 is} hydrogen or a C 1 -C ₃ -alkoxy radical which contains one or more hydroxyl group (s), thiol group (s) or Amino group (s) and / or halogen atom (s) may be substituted,

R ^{4 is} hydrogen or a C 1 -C ₃ -alkoxy radical which may be substituted by one or more hydroxyl group (s), thiol group (s) or amino group (s) and / or halogen atom (s),

can mean

The present invention particularly preferably relates to matrix materials according to the general formula I.

T - (C = O) O - B - D (I) in which

T is a crosslinked polymer consisting of units of methacrylic acid esters

B is a covalent bond or a methylene group

D is -CH ₂ -CH (OH) -CH ₂ OH, -CH ₂ -C (CH ₂ OH) ₃ , -CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -OH, or a group

-CH (XR ¹ ) -CH ₂ (YR ² ),

in the

X and Y oxygen, R ¹ and R ^{2 are} independently hydrogen, -CH (OH) -CH ₂ OH, -CH ₂ -CH (OH) -CH ₂ OH ₅ -CH ₂ -C (CH ₂ OH) ₃ , -CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -OH,

can mean.

The present invention very particularly preferably relates to matrix materials according to the general formula I.

T - (C = O) O - B - D (I) in which

T is a polymer cross-linked with ethylene glycol dimethacrylate and consists of units of preferably one or more methacrylic acid ester (s),

B is a covalent bond or a methylene group.

D - CH ₂ -CH ₂ OH, -CH ₂ -CH (OH) -CH ₂ OH, -CH ₂ -C (CH ₂ OH) ₃ , -CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -OH or a group -CH (XR ¹ ) -CH ₂ (YR ² ),

in the

X and Y oxygen,

R ¹ and R ^{2 are} independently hydrogen, -CH (OH) -CH ₂ OH, -CH ₂ -CH (OH) -CH ₂ OH ₅ -CH ₂ -C (CH ₂ OH) ₃ , -CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -OH,

can mean. More particularly, the present invention relates to matrices comprising any of the above-described matrix materials bonded to a material having a permanent or temporary magnetic moment.

The present invention preferably relates to the matrices described above, which have as a permanent or temporary magnetic moment having materials which are ferromagnetic or ferromagnetic or para or superparamagnetic.

The present invention preferably relates to the above-described matrices comprising a matrix material which is in the form of beads.

The present invention furthermore preferably relates to the above-described matrices comprising a matrix material in which the matrix is in the form of a composite.

In addition, the present invention relates to processes for the preparation of the above-described matrix materials, which comprises the following steps:

a) polymerizing the support material and the monomers carrying the epoxy partial structure, if appropriate in the presence of a material which has a permanent or temporary magnetic moment, are polymerized, and

b) subjecting the resulting polymers to hydrolysis, alcoholysis, aminolysis or reaction with a thiol, and

c) the derivatized polymers are isolated.

The present invention further relates to processes for the preparation of the above-described matrix materials, comprising the following steps:

a) the monomers constituting the support material and the monomers carrying the ester partial structure, if appropriate in the presence of a crosslinking agent and / or a Material that has a permanent or temporary magnetic moment, is polymerized and

b) the resulting polymers are transesterified with an alcohol and

c) the derivatized polymers are isolated.

The present invention also relates to the use of matrix materials according to the general formula I.

T - (C = O) O - B - D (I) in which

B is a covalent bond or an alkylene group having 1 to 6 carbon atoms

D is a branched or unbranched C ₂ -C _2O -AI kyrest or C ₃ -C ₂ -

-CH (XR ¹ ) - CH ₂ (YR ² ),

in the X and Y independently of one another oxygen, sulfur or -NR ³ -

and

R ¹ and R ² are independently hydrogen or a branched or unbranched Ci -C _2O -AI kyrest or C ₃ - Ci2-cycloalkyl radical whose carbon chain optionally interrupted by one to 10 oxygen, sulfur or divalent amino group (s) -NR ⁴ - , the same or different, may be interrupted and which may be interrupted by one or more hydroxyl group (s), thiol group (s) or amino group (s), aldehyde group (s), carboxylic acid amide group (s), sulfonamide group (s) and / or halogen atom (s) may be substituted

can mean for purifying or isolating biomolecules.

The present invention preferably relates to the use of matrix materials according to the general formula I. T - (C = O) O - B - D (I) in which

T is a polymer or copolymer comprising units of

B is a covalent bond or an alkylene group having 1 to 3 carbon atoms

D is a branched or unbranched C ₂ -C ₀ -alkyl radical or C ₃ -C ₀ -

-CH (XR ¹ ) - CH ₂ (YR ² ),

in the

X and Y are independently oxygen, sulfur or -NR ³ -,

R ¹ and R ² are independently hydrogen or a branched or unbranched _O -alkyl radical or C ₄ CRCI - cycloalkyl radical -C whose carbon chain optionally substituted by one or up to three oxygen, sulfur or bivalent amino group (s) -NR ⁴ -, the same or different, may be interrupted and the one or more hydroxyl group (s), thiol group (s) or amino group (s), aldehyde group (s), Carboxamide group (s), sulfonamide group (s) and / or halogen atom (s) may be substituted,

can mean for purifying or isolating biomolecules.

The present invention more preferably relates to the use of matrix materials according to the general formula I.

T - (C = O) O - B - D (I) in which

T is a polymer or copolymer comprising units of acrylic acid and / or methacrylic acid and / or their esters

B is a covalent bond or a methylene group

D is a branched or unbranched C ₂ -C ₈ -alkyl radical or C ₃ -C ₈ -

Cycloalkyrest, whose carbon chain may optionally be interrupted by one to two oxygen, sulfur or bivalent amino group (s) -NR ⁴ -, the same or different, and having one or more hydroxyl group (s), thiol group (s) or amino group (n) Aldehyde group (s), carboxylic acid amide group (s), sulfonamide group (s) and / or halogen atom (s), or a group

-CH (XR ¹ ) -CH ₂ (YR ² ),

in the

X and Y are independently oxygen, sulfur or -NR ³ -,

R ¹ and R ² independently of one another denote hydrogen or a branched or unbranched C ₁ -C ₆ -alkest or C ₄ -C ₆ -cycloalkyl radical whose carbon chain may optionally be replaced by one or with up to two oxygen, sulfur or bivalent amino group (s) - NR ⁴ -, same or different, and may be interrupted by one or more hydroxyl group (s), thiol group (s) or amino group (s), aldehyde group (s), carboxylic acid amide group (s), sulfonamide group (s) and / or halogen atom ( en) can be substituted,

R ^{3 is} hydrogen or a C 1 -C ₃ -alkoxy radical which may be substituted by one or more hydroxyl group (s), thiol group (s) or amino group (s) and / or halogen atom (s),

R ^{4 is} hydrogen or a C 1 -C ₃ -alkoxy which may be substituted by one or more hydroxyl group (s), thiol group (s) or amino group (s) and / or halogen atom (s),

can mean for purifying or isolating biomolecules. The present invention particularly preferably the use of matrix materials according to the general formula I.

T - (C = O) O - B - D (I) in which

T is a crosslinked polymer consisting of units of methacrylic acid esters

B is a covalent bond or a methylene group

-CH (XR ¹ ) -CH ₂ (YR ² ),

in the

X and Y oxygen,

can mean for purifying or isolating biomolecules.

The present invention very particularly preferably the use of matrix materials according to the general formula I. T - (C = O) O - B - D (I) in which

B is a covalent bond or a methylene group.

in the

X and Y oxygen,

can mean for the purification or isolation of biomolecules.

More particularly, the present invention relates to the uses of the inventive matrices for purifying biomolecules described above, wherein the biomolecule embodies one or more species of nucleic acid (s).

In addition, the present invention relates to a method for isolating biomolecules with one of the matrices according to the invention, comprising the following steps: (a) providing the matrix;

(c) incubating the matrix and the mixture, wherein the biomolecule or preferably the desired nucleic acid is immobilized on the matrix;

(e) combining the matrix with the biomolecule or with the immobilized nucleic acid (s) with an elution solution, wherein the desired biomolecule (s) is desorbed from the matrix.

In particular, the present invention relates to a method of isolating biomolecules with one of the matrices of the invention, wherein the biomolecule is one or more species of nucleic acid (s).

In addition, the present invention relates to nucleic acids which are bound in the form of a complex with the matrix material according to the invention.

In addition, the present invention relates to a microfluidic system comprising a matrix material or the use of the matrix material according to the invention in or in conjunction with a microfluidic system.

Claims

claims

1 . Matrix material according to the general formula I.

T - (C = O) O - B - D (I) in which

B is a covalent bond or an alkylene group having 1 to 6 carbon atoms

D is a branched or unbranched C ₂ -C _2O -AI kyrest or C ₃ -C ₂ -

-CH (XR ¹ ) - CH ₂ (YR ² ),

in the

X and Y independently of one another oxygen, sulfur or -NR ³ -

and

R ¹ and R ² are independently hydrogen or a branched or unbranched CrC kyrest ₂₀ -AI or C ₃ - C ₂ -Cycloalkyrest whose carbon chain optionally interrupted by one to 10 oxygen, sulfur or divalent amino group (s) -NR ⁴ -, same or may be different, and may be substituted with one or more hydroxyl group (s), thiol group (s) or amino group (s), aldehyde group (s), carboxylic acid amide group (s), sulfonamide group (s) and / or halogen atom (s) .

can mean

2. matrix material according to claim 1, characterized in that

T is a polymer or copolymer comprising units of

B is a covalent bond or an alkylene group having 1 to 3 carbon atoms

D is a branched or unbranched C ₂ -C ₀ -alkyl radical or C ₃ -C ₀ -

-CH (XR ¹ ) - CH ₂ (YR ² ),

in the

X and Y are independently oxygen, sulfur or -NR ³ -,

R ¹ and R ² are independently hydrogen or a branched or unbranched CRCI _O -alkyl radical or C ₄ - Cio-cycloalkyl radical whose carbon chain optionally substituted by one or up to three oxygen, sulfur or divalent amino group (s) -NR ⁴ -, the same or different, and may be interrupted by one or more hydroxyl group (s), thiol group (s) or amino group (s), aldehyde group (s), carboxylic acid amide group (s), sulfonamide group (s) and / or halogen atom (s) ), R ^{3 is} hydrogen or a C 1 -C ₆ -alkoxy which may be substituted by one or more hydroxyl group (s), thiol group (s) or amino group (s) and / or halogen atom (s),

R ^{4 is} hydrogen or a C ₁ -C ₆ -alkoxy which may be substituted by one or more hydroxyl group (s), thiol group (s) or amino group (s) and / or halogen atom (s),

can mean

3. matrix material according to claim 2, characterized in that

B is a covalent bond or a methylene group

D is a branched or unbranched C ₂ -C ₈ -alkyl radical or C ₃ -C ₈ -

-CH (XR ¹ ) -CH ₂ (YR ² ), in the

X and Y are independently oxygen, sulfur or -NR ³ -,

R ^{3 is} hydrogen or a C 1 -C ₃ -alkoxy which may be substituted by one or more hydroxyl group (s), thiol group (s) or amino group (s) and / or halogen atom (s),

can mean

4. matrix material according to claim 3, characterized in that

T is a crosslinked polymer consisting of units of methacrylic acid esters

B is a covalent bond or a methylene group

-CH (XR ¹ ) -CH ₂ (YR ² ),

in the

X and Y oxygen,

can mean.

5. matrix material according to claim 4, characterized in that

B is a covalent bond or a methylene group.

in the

X and Y oxygen,

can mean.

6. matrix containing a matrix material according to one of claims 1 to 5, characterized in that the matrix material is connected to a material having a permanent or temporary magnetic moment.

7. matrix containing a matrix material according to claim 6, characterized in that the material having a permanent or temporary magnetic moment is ferromagnetic or ferromagnetic or para or superparamagnetic.

8. matrix containing a matrix material according to any one of claims 1 to 5, characterized in that it is in the form of beads.

9. matrix containing a matrix material according to any one of claims 1 to 5, characterized in that the matrix is in the form of a composite.

10. A process for the preparation of matrix materials according to any one of claims 1-5, characterized in that a) polymerizing the support material and the monomers carrying the epoxy partial structure, if appropriate in the presence of a material which has a permanent or temporary magnetic moment, are polymerized, and

c) the derivatized polymers are isolated.

1 1. Process for the preparation of matrix materials according to one of claims 1-5, characterized in that

a) which polymerizing the support material and carrying the ester partial structure monomers, if necessary in the presence of a crosslinking agent and / or a material having a permanent or temporary magnetic moment, are polymerized, and

b) the resulting polymers are transesterified with an alcohol and

c) the derivatized polymers are isolated.

12. Use of a matrix material according to the general formula I.

T - (C = O) O - B - D (I) in which

B is a covalent bond or an alkylene group having 1 to 6 carbon atoms a branched or unbranched C ₂ -C _2O -AI kyrest or C ₃ -C ₂ - cycloalkyl radical whose carbon chain optionally interrupted by one to 10 oxygen, sulfur or divalent amino group (s) -NR ⁴ -, identical or different, interrupted and which may be substituted with one or more hydroxyl group (s), thiol group (s) or amino group (s), aldehyde group (s), carboxylic acid amide group (s), sulfonamide group (s) and / or halogen atom (s), or a group

-CH (XR ¹ ) - CH ₂ (YR ² ),

in the

X and Y independently of one another oxygen, sulfur or -NR ³ -

and

R ¹ and R ² are independently hydrogen or a branched or unbranched CrC kyrest ₂₀ -AI or C ₃ - C ₂ -Cycloalkyrest whose carbon chain optionally interrupted by one to 10 oxygen, sulfur or divalent amino group (s) -NR ⁴ -, may be the same or different, interrupted and substituted with one or more hydroxyl group (s), thiol group (s) or amino group (s), aldehyde group (s), carboxylic acid amide group (s), sulfonamide group (s) and / or halogen atom (s) can be,

R ^{3 is} hydrogen or a C 1 -C ₆ -alkoxy which may be substituted by one or more hydroxyl group (s), thiol group (s) or amino group (s) and / or halogen atom (s), R ^{4 is} hydrogen or a C 1 -C ₆ -alkoxy radical which may be substituted by one or more hydroxyl group (s), thiol group (s) or amino group (s) and / or halogen atom (s),

can mean

for cleaning or isolating biomolecules.

13. Use according to claim 1 2, characterized in that

T is a polymer or copolymer comprising units of

B is a covalent bond or an alkylene group having 1 to 3 carbon atoms

D is a branched or unbranched C ₂ -C ₀ -alkyl radical or C ₃ -C ₀ -

-CH (XR ¹ ) - CH ₂ (YR ² ),

in the

can mean.

14. Use according to claim 13, characterized in that

B is a covalent bond or a methylene group

D is a branched or unbranched C ₂ -C ₈ -alkyl radical or C ₃ -C ₈ -

Cycloalkyrest, whose carbon chain optionally by a up to two oxygen, sulfur or bivalent amino group (s) -NR ⁴ -, which may be identical or different, and which is interrupted by one or more hydroxyl group (s), thiol group (s) or amino group (s), aldehyde group (s), Carboxylic acid amide group (s), sulfonamide group (s) and / or halogen atom (s) may be substituted or a group

-CH (XR ¹ ) - CH ₂ (YR ² ),

in the

X and Y are independently oxygen, sulfur or -NR ³ -,

R ^{4 is} hydrogen or a C 1 -C ₃ -alkoxy radical which may be substituted by one or more hydroxyl group (s), thiol group (s) or amino group (s) and / or halogen atom (s), can mean.

15. Use according to claim 14, characterized in that

T is a crosslinked polymer consisting of units of methacrylic acid esters

B is a covalent bond or a methylene group

-CH (XR ¹ ) -CH ₂ (YR ² ),

in the

X and Y oxygen,

can mean.

16. Use according to claim 15, characterized in that

B is a covalent bond or a methylene group.

in the

X and Y oxygen,

can mean.

17. Use according to any one of claims 12 to 16, characterized in that the biomolecule embodies one or more species of nucleic acid (s).

18. A method for isolating biomolecules with a matrix according to any one of claims 1-5, characterized in that it comprises the following steps:

(a) providing the matrix; (b) combining the matrix with a mixture containing the biomolecule to be isolated in addition to at least one contaminant;

(c) incubating the matrix and the mixture, wherein the biomolecule is immobilized on the matrix;

(d) separating the matrix with the biomolecule or mixture;

(e) combining the matrix with the biomolecule or with an elution solution, wherein the desired biomolecule (s) is desorbed from the matrix.

19. The method according to claim 18, characterized in that the biomolecule represents one or more species of nucleic acid (s).

20. Nucleic acid characterized in that it is bound in the form of a complex with the matrix material according to any one of claims 1-9.

21. Microfluidic system comprising a matrix material according to any one of claims 1-9.