FI115294B - Handling macroparticles useful as solid phase to bind e.g. biomolecules involves performing at least two treatment steps in same vessel without moving the particles to another vessel - Google Patents

Abstract

Handling macroparticles (22), comprising performing at least two treatment steps in a same vessel (26) without moving the particles to another vessel, is new. An independent claim is included for a device for handling microparticles.

Description

115294 MAGNETIC TRANSMISSION, MICROPARTIC TRANSMITTER AND REACTION UNIT

FIELD OF THE INVENTION

The invention relates to a magnetic transfer method.

BACKGROUND OF THE INVENTION

Magnetic transfer means any activity related to the motion of particles by means of magnetism, such as sorting, collecting, moving, mixing or dispensing particles in or between liquids.

Particles, micro particles or magnetic particles

15 magnetic particles) refers to any small particle having a diameter in the predominantly micrometer range that can be moved by magnetism. There are many different types of known magnetically transferable particles and the applications in which they are used also vary widely. For example, the particles used in microbiology generally have a size of 0.01 to 100 µm, most commonly 0.05 to 10 µm. Known such particles 20 are, for example, particles containing ferromagnetic, paramagnetic or supramagnetic material. The particles can also be magnetic in themselves, so they can be moved by any ferromagnetic body.

; : The microparticle processing device has a magnetism utilizing member, •: * · · 25, hereinafter referred to as a magnet. It may be a permanent magnet or: v. An electromagnet attracting ferromagnetic particles, or ferromagnetic, · *. a piece that is not magnetic in itself, but still attracts magnetic particles.

* *, The magnet is usually most preferably a circular rod magnet. It can also be other · ;;; 30 shaped rod. However, the magnet need not be a rod at all. It can also be * i 'short and wide, or any shape. The magnet may also be composed of *: ··: one or more pieces, such as magnets or ferromagnetic •: · · · pieces.

:, '35 There must be a cover on the magnet that protects the magnet from various harmful - "· conditions and allows the microparticles to be manipulated, such as binding and release.

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The structure of the shield can vary greatly, for example in the form of a thin film of flexible or stretchable material or even a cup of hard plastic.

In general, microparticles are used as a solid phase to bind 5 different biomolecules, cellular organelles, bacteria, or cells. For example, enzymes can also be immobilized on the surface of the microparticles, thus further utilizing the enzyme treatments. Most of the so-called magnetic nanoparticles (<50 nm) are not suitable for treatment with conventional permanent magnets or electromagnets, but require the use of a particularly strong magnetic gradient as disclosed in EP 10 0842704 (Miltenyi Biotec). Conventional permanent and electromagnets can usually handle magnetic particles, such as microparticles, having a diameter of about 0.1 µm or larger. The viscosity of the sample can also make it difficult to pick up the particles significantly. The particles to be collected may initially be suspended in a large volume of liquid from which to bind the test substance or, for example, cells to the surface of the particles. It is particularly important to be able to use high initial volumes in applications from which low-volume components are to be isolated for analysis. For example, efficient enrichment of pathogenic bacteria from a large sample volume to a small sample volume is critical because it directly affects the sensitivity and assay of the assay. At present, there is no efficient enough way to make microparticles 20 concentrations from a large volume to a small volume. It would be advantageous if the performance described above is as simple and effective as possible.

: BACKGROUND OF THE INVENTION

*; Magnetically processed microparticles have been used since the 1970s. These 25 technologies became very popular among others in immunoassays. The use of microparticles in immunoassays to separate the bound antigen-antibody complex from the free fraction provided a significant advantage especially in the reaction rate.

The main development in microparticle utilization in recent years is Γ · '; occurred in the fields of molecular biology, microbiology and cell biology.

30,;. In the conventional method, magnetic particles in the reaction solution, such as "microparticles, are trapped by a magnet outside the vessel at a specific point on the inner wall of the tube. The solution is then carefully cautiously removed from the magnetic particles: V. I drive.

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Another approach uses magnets to actively transfer microparticles. The magnet is pushed into the solution containing the microparticles, whereby the magnet attracts the microparticles in the solution and forms a solid precipitate. The magnet and microparticles can then be removed from the liquid.

The magnet and its particles can then be immersed in the liquid in the second test tube, where the microparticles can be detached from the magnet. In this method, solution handling, pipetting and aspiration phases are minimized to the extreme.

US Patent 2,517,325 (Lamb) discloses a solution for picking metal objects by means of a magnet. The publication describes a long rod magnet that is moved inside an iron tube. The poles of the rod magnet are at opposite ends of the longitudinal axis of the physical magnet. By moving the magnet inwards in the iron tube, the magnetic field can be reduced. Similarly, by moving the magnet out of the iron tube, the magnetic field is enhanced.

The publication describes a solution for collecting metal objects on the tip of a magnetic unit. The publication also describes a solid plastic shield for protecting the magnet.

US 2,970,002 (Laviano) describes a solution for collecting metal objects 20 from liquids by means of a magnet. The publication describes a long permanent magnet which | . \ Collect particles at the tip of the magnetic unit. The magnet is attached to a metal bar and * ·· ·; , ·. protected by a separate plastic cover. The publication discloses the combined use of moving a permanent magnet and a plastic cover to protect the * · j magnet. This publication describes the collection of metal objects at the tip of a magnetic unit and the release of metal objects.

The Mlit 25 is protected by a special plastic cover design.

* »·

US 3,985,649 (Eddelman), US 4,272,510 (Smith et al), US 4,649,116 (Daty et al), US 4,751,053 (Dodin et al) and US 5,567,326 (Ekenberg et al.) Describe solutions, where all magnets collect material to be magnetized directly from 30 solutions. What these publications also have in common is that the magnets are not protected by separate plastic protectors. These solutions require washing of the magnetic tip before the next ''. treatment of the sample to eliminate contamination and carry-over effects.

U.S. Patent No. 5,288,119 to Crawford, Jr. et al. Describes a solution for collecting metal objects by means of a magnet. The magnet of the device of the publication is not protected by a special shield and is not suitable for picking metal objects from liquids.

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The publication describes a solution for picking up larger metal objects. The publication discloses a long rod magnet which is moved within a non-magnetic tube. A special feature of this tube is that it acts as a magnetic blocking agent, even though it is not magnetic. The publication discloses, for example, bismuth or lead or a mixture thereof as alternative materials for this purpose. The magnet of the solution is not specially protected and is not suitable for picking metal objects from liquids.

WO 87/05536 (Schroeder) describes the use of an io permanent magnet movable within a plastic shield for collecting ferromagnetic material from a solution containing them. When the magnet is in the lower position, the ferromagnetic material collects at the tip of the magnet unit. The publication describes the transfer of the ferro-magnetic material thus collected to a solution in the second vessel and the release of the material from the tip portion there. The release of the ferromagnetic material is described as being accomplished by the design of a plastic shield that prevents the material from moving upwardly as the magnet is moved.

US 5,837,144 (Bienhaus et al.) Describes a method for collecting microparticles by means of a special magnet with a plastic shield. This publication 20 describes the binding of microparticles from a solution which is discharged from the vessel by various arrangements. By moving the magnet, the microparticles can be released = = ·: v. From the protective film.

'! U.S. Patent Nos. 5,942,124 (Tuunanen), US 6,020,211 (Tuunanen), US 6,040,192, 25 (Tuunanen), US 6,065,605 (Korpela et al.) And US 6,207,463 (Tuunanen), and &gt; U.S. Patent Application Publication No. 20010022948 (Tuunanen) also describes devices with a plastic shield for collecting microparticles from a solution and transferring it to another solution. These publications mainly describe solutions for treating microparticles in very small volumes. US 5,942,124: ': 30 (Tuna) discloses a device for concentrating microparticles at the very tip of the magnetic unit. US 6,020,211 (Tuna) is described in the previous [. The apparatus disclosed in the publication for use with a large so-called conventional magnet for transferring the collected microparticles into smaller containers. US 6,040,192:, · ', · (Tuunanen) describes an automated method for the use of microparticles in specific': 35 assays and small volume processing. US 6,065,605 (Korpela et al.) Continues to apply the solution described in US 5,942,124 (Tuunanen) to the treatment of larger volumes. A method is now described in which 5,152,954 microparticles are first collected by a special large magnet containing magnetic unit. Thereafter, the magnetic unit described in US 5,942,124 (Tuunanen) is used to move the microparticle pellet forward into smaller containers. US 6,207,463 (Tuna) also applies the above-described magnetic unit capable of collecting microparticles at the very tip of the device. US 20010022948 (Tuunanen) also describes the treatment of a very small amount of microparticles in special containers designed for it.

US 6,403,038 (Heermann) describes a device having a plastic shield and a permanent magnet attached to 10 special bars. The microparticles are collected at the tip of the plastic shield and the method is specifically designed to handle small volumes. The rod has a special protruding part that keeps the magnet and the rod in place in the protective tube.

EP 1058851 (Ripple) and WO 01/60967 (Ripple) 15 describe devices having an elastic elastomeric protective film. In these solutions, microparticles are collected on the surface of a stretchable protective film, from where they can be further transferred to another container. The magnet's protective film is made of stretchable material, so that the film is stretched as thin as possible. This provides a minimum distance from the magnet to the liquid.

20

US 5,610,077 (Davis et al.) Describes a particular inner tube and outer tube; '·. in conjunction with specific immunoassays. This publication describes immunoassays performed in a special tube or in a well of a * * t '. * Microtiter plate, or microplate, with a small volume of fluid, by means of an internal tube arrangement. With the tube arrangement in question,,, a small »» »', t in the test tube or microplate well can be obtained; * increases the volume of the liquid to increase the reactive surface of the tube * · '· ·' and solution efficient mixing. Microparticles and concentration from high volume to low volume are not mentioned in the publication.

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None of the above-described patents describe a method that could efficiently collect microparticles from very large volumes of liquid and release the collected particles to a smaller volume of liquid. In particular, no realistic way of collecting a large number of microparticles from a large volume of liquid has been described. Rather, the abovementioned publications of v 'describe the treatment of smaller liquid volumes, such as 5-10 ml 35, and very small liquid volumes. If it is desired to bind proteins, peptides, nucleic acids, cells, bacteria, viruses, or other components from a large volume to the surface of microparticles, there are certain basic conditions for the optimal particle number to be used. Depending on the microparticles used, the preferred amount of particles per milliliter of liquid to be isolated may be, for example, at least 107 microparticles, e.g., 1 to 5 µm in diameter. The number of particles required will continue to increase if the desired, very small number of components are to be bound as reliably as possible within a given unit volume.

In particular, the microparticles described in US 5,942,124 (Tuunanen), US 6,020,211 (Tuunanen), US 6,065,605 (Korpela et al), and US 6,207,463 (Tuunanen) and EP 0 787 296 (Tuunanen) are intended to be collected from a large very sharp and narrow to the small tip of the rod, is impractical.

A large number of microparticles cannot be shifted to a small volume around a small point because the physical dimensions of the pellet formed by the microparticle mass increase rapidly with the volume of liquid to be treated. The large mass of microparticles must be collected either in a large area or in a special well.

PURPOSE OF THE INVENTION

It is an object of the present invention to provide a method and apparatus which do not have the drawbacks set forth above. The magnetic transfer method according to the invention is characterized in that:

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• »!! . The invention relates specifically to the active collection and transfer of microparticles from one fluid to another. In particular, the method can be used in automated equipment, where • various microparticle transfers, washes and incubations can be performed.

'* 25 It is possible to combine units with the purpose of * »·:.' With automated equipment, for example for the detection of PCR reactions or various labels.

A TRANSMISSION DEVICE OF THE INVENTION

The invention also relates to a microparticle transfer device.

30, The essential technical feature of the device according to the invention is that the intensity and orientation of the magnetic field relative to the protective film surrounding the magnet can be controlled. This can be accomplished by moving the magnet in the ferromagnetic tube V: so that it can be completely inside the tube, whereby the power of the magnet is negligible or non-existent, or it can be partially or totally outside the tube, with the power and collecting surface parts. Combining these features of 115294 7 into magnetic containers for transferring magnetic particles provides a highly efficient collection and concentration process.

The tube may be made of iron or other suitable material having magnetic properties suitable to prevent magnetic flux from passing through the tube. The power of the magnet can be controlled by changing the position of the magnet relative to the ferromagnetic tube so that part of the magnet is inside the tube. Alternatively, the magnet may be held in place and the ferromagnetic tube moved relative to the magnet. The magnet is mounted on a rod, ferromagnetic or non-ferromagnetic, which allows the magnet to be moved within the ferromagnetic tube.

The properties and advantages of the ferromagnetic tube mentioned in the invention are at least the following: 1. The tube protects the magnet and its coating against mechanical stress 15 2. The tube reinforces the structure of the magnetic bar and in particular the connection between the tube and the movable pin. sensitive devices, especially when the magnet is inside the tube 20 5. The tube can be stretched and / or shaped with a stretchable protective film>> '· * The magnet may take the form of a round rod or pin, for example, but it may also be t »! ! other shape. The magnet excitation axis may also vary. The magnetization axis can be either longitudinally parallel to the longitudinal axis of the bar and * · · · · [25 magnet poles are at the ends of the bar. In this case, the excitation is in the same direction as the · · 'ferromagnetic tube, i.e. the direction of motion of the magnet or tube.

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However, the magnet excitation axis can also be transverse to: '· perpendicular to both the ferromagnetic tube and the longitudinal axis of the rod-shaped magnet:': 30. The magnetization direction is then perpendicular to the direction of motion of the magnet or tube.

On the other hand, a magnet may also consist of several different magnets, which may be the same or different, and which may be attached to one another by magnetic force: '·,: 35 or by a material ferromagnetic or non-ferromagnetic.

The magnet may also be a combination of magnetic and ferromagnetic material. The magnet may also be either a permanent magnet or an electromagnet.

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The magnetic arrangement of the invention, the protective film and the containers used can be very effective in treating microparticles in both large and small volumes of liquid. The concentration of the microparticles just near the tip portion 5 of the magnetic unit allows both concentration in large volumes and handling of the microparticles in small volumes. Thus, the invention describes a universal solution for applications with microparticles both on a large and small scale.

The invention provides a solution that is optimal for use on a large scale to collect and transfer microparticles from both large and small liquid volumes. In particular, the invention assists in collecting particles from high fluid volumes and releasing them into low fluid volumes.

15 The invention provides a special offering an adequate support to the plastic cover or the outer side of the elastomer collected mikropartikkelimassan design cheaper and more reliable protection around the collection. Special design refers, for example, to grooves, pits and / or bumps of various sizes and depths. As these designs roll over, the microparticle pellet receives special support for protection when moving the magnetic unit and against fluid flows. The effect of viscous samples is very significant:. *. which in the worst case means that the microparticles do not stick to the side of the guard • X but remain in solution. For handling large volumes, the above design has! naturally a major benefit to collection security.

The device and method described in the invention can be implemented in the treatment of very large volumes and, on the other hand, can be applied in small volumes.

...: A particularly effective method is to optimize the magnetic unit, the vessels used with it, and the fluid volumes. In particular, using a volume displaced by a magnetic unit to adjust the height of the liquid surface is a "*: 30 highly effective method in the concentration step. For the first time, the apparatus and method are described which microparticle collection area, intensity, and physical location of microparticles can be adjusted.

The invention describes a device and method for collecting microparticles in a plurality of 35 applications. A central technical solution of the invention is the ability to control the force and orientation of the magnetic field by means of a ferromagnetic tube on the surrounding protective film around which the microparticles are collected. The magnet can be moved in and out of 115,294 g of ferromagnetic tube, thereby changing the magnetic field of the magnet. When the magnet is outdoors, the protective film is exposed to a magnetic field the same size as the magnet outside the ferromagnetic tube. The microparticles can then be collected outside the protective film. When the magnet is moved 5 completely inside the ferromagnetic tube, there is no significant magnetic field externally applied. In this case, the microparticles do not accumulate around the protective film but remain in solution. The tube can be fixed or adjustable to achieve the best collection efficiency.

The method and apparatus of the invention provide the following solutions and features: 1. Collecting microparticles from a large volume of liquid.

2. Collecting a large number of microparticles.

3. Using the same device to collect small amounts of liquid and small amounts of microparticles.

4. Collecting microparticles only at one end of the magnet or over the entire surface of the magnet.

5. Collecting microparticles using a rigid plastic shield.

6. Collecting microparticles using an elastic elastomeric shield.

20 7. Various movements, such as the magnet or the sleeve around it, ·. utilization.

v! 8. Using different containers for concentration.

! 9. Release of microparticles into a small amount of liquid.

•; 10. Use of different magnets to achieve optimum microparticle collection geometry MM · 25.

> »* · '11. Effective mixing.

• · · :: 12. Sealing a test tube or microplate well or well with a protective film.

The microparticles may contain affinity ligands, enzymes, antibodies, bacteria, cells or organelles. Binding of the desired components can also be obtained. by selecting the surface properties of the microparticles used and the composition of the buffers suitably advantageous to bind the desired components from the samples. Examples include 'ion-exchange, hydrophobic and reverse-phase chromatography. In these, for example, binding and release of proteins from the surface of microparticles is conveniently performed; *! 35 selected buffers and solutions. Very important factors are, for example, salt content and pH.

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The affinity ligand may be, for example, a single or double stranded nucleotide sequence such as DNA (Deoxyribonucleic Acid), RNA, mRNA or cDNA (Complementary DNA), or PNA (Peptide Nucleic Acid), protein, peptide, polysaccharide, oligosaccharide, small molecule compound. The affinity ligand can also be any of the following: Ovomucoid, ProteinA, Aminophenyl boronic acid, Procion red, Phosphoryl ethanolamine, Protein G, Phenyl alanine, Proteamine, Pepstatin, Dextran sulfate, EDTA (Ethylenediaminetetraacetic Acid), PEG (Polyethylene) -glucosamine, Gelatin, Glutathione, Heparin, Iminodiacetic add, NTA (Nitrilotriacetic Add), Lentil Lectin, Lysine, NAD (Nicotinamide Adenine Dinudeotide), Aminobenzamidine, Acriflavine, AMP, 10 Aprotinin, Avidin, Streptavidin, Bovine Serum (Bovine) Biotin, Concanavalin A (ConA) and Cibacron Blue.

Immobilization of the enzyme or affinity ligand to the microparticles means that the enzyme or ligand is attached to the surface of the particles or trapped within 15 "chipped" particles, but in contact with the surrounding solution.

The attachment of the enzyme or affinity ligand to the microparticles can be accomplished by a covalent bond, for example by amino or hydroxy groups on the support. Alternatively, the binding may be effected by means of a bioaffinity pair, for example a biotin / streptavidin pair. According to one method, the enzyme to be immobilized is produced by recombinant DNA technology, for example in Escherichia coli, and the enzyme has been subjected to a specific affinity tail. This affinity tail binds to microparticles with »·:; a component which is highly bound to the affinity tail and is appropriately attached.

The affinity tail can be a small molecule compound or a protein. Such an arrangement could efficiently utilize microparticles of the desired enzyme to purify the desired enzyme, and at the same time the microparticle-bound enzyme would be pre-immobilized on the microparticle surface for use in the process of the invention.

♦ »30 ,. The attachment of the enzyme or affinity ligand to the microparticles may also be nonspecific, non-covalent, such as adsorption.

The invention relates to a device and a method for collecting microparticles from containers of very different sizes and transferring microparticles from one container to another. In particular, the invention describes an apparatus for collecting microparticles from a large volume and concentrating them in a smaller volume. The term "microparticle", as used herein, refers to particles having a size preferably from 0.10 to 100 µm. The microparticle may also be a much larger particle, for example, a particle of several millimeters in diameter. In the invention, microparticles are magnetic such as para, superpara , or magnetizable material, or the microparticles are attached to a magnetic or magnetizable body and that the microparticles to which, for example, affinity groups or enzymes may be attached, are captured by a magnetic unit embedded in the first vessel, transferred to the second vessel, Alternatively, the microparticles do not need to be specifically removed from the magnetic unit.

The magnet by which the particles are trapped can be either a permanent magnet or an electromagnet. The shape of the magnets may vary depending on the application. The magnetic field in the magnets may be different: a magnet magnetized in the longitudinal direction, 15 in the same direction as the diameter of the magnet magnetized, or several magnet poles in the same magnetic body. The individual magnets may also be connected to one another or by means of suitable ferromagnetic or non-ferromagnetic spacers.

The protective film may be of non-stretchable material such as polypropylene, polystyrene, polycarbonate, polysulfone and polyethylene. The protective film may also be non -:. ferromagnetic metal or ferromagnetic metal. The protective film may also be stretchable, ·! elastomeric material such as silicone rubber, fluoroelastomer, polychloroprene, polyurethane or chlorosulfonated polyethylene. The protective film may also be treated with special agents to alter the properties of the protective film. The protective film 25 may thus be coated with, for example, Teflon (PTFE, Polytetrafluoroethylene). It is particularly important to be able to select the protective material and any further treatment so that the end result:.,.: Allows the operation according to the invention even with very strong or corrosive chemicals. The protective film may also be shaped to allow multiple; protection of separate magnetic units, for example on 8.12 or 96-channel devices.

'' ': 30 The shape of the protective film may be either tubular, plate-shaped or irregularly shaped.

There are especially many possibilities when using an elastomeric protective film, because then the inner magnet and the ferromagnetic tube can also shape the protective film.

;. · A preferred alternative to a protective film is a flat or sheet-like, stretchable material: '; 35. Such a protective film may be a single stretchable film in a special frame. The purpose of the frame is to facilitate the use of the protective film and to provide the film with stretching properties. Another alternative is a roller-like 115294 12 embodiment, whereby the change of the protective film can be done simply by rolling the new protective film off the roll. This option may also include the use of a frame, special support or bracket when stretching the protective film during actual use. The use of such a single sheet protective film is a highly recommended option 5 when it comes to reducing material consumption during insulation and cleaning operations. It is also economically cheaper to use a sheet-like protective film than shaped and large-size protective films made with mold tools.

10 Applying a sheet-like protective film to an automatic device is a very simple and effective option. When using a sheet-like protective film, the initial stretching of the ferromagnetic sleeve can be performed in the first step. At this point, the magnet is still inside the ferromagnetic sleeve and the microparticles outside the protective film are not exposed to the magnetic field. While the protective film is still kept stretched, the magnet can conveniently be expelled from inside the ferromagnetic sleeve. In this case, the magnet still stretches the shielding membrane. By moving the magnet inside or out of the sleeve, the solution in the test tube is stirred with the aid of a magnet. The agitation can also be performed by moving the ferromagnetic sleeve 20 up and down.

> ♦ 1 · V The particularly preferred embodiment shown above is for microparticles processing! in small containers, such as microplates with 96 or 384 wells. Presented '; the solution and solution of microparticles are advantageous because the whole device does not have to be moved * * * · · 25. The mixing is done by simply moving the magnet and / or ferromagnetic sleeve. The particularly optimal solution presented is because the process is not needed

Ml \; traditional shakers at all. After all, it is a known fact that conventional shakers are not capable of effectively mixing small amounts of solution, and in particular of holding microparticles in solution. A major problem with known devices is the rapid sedimentation of microparticles: * * ': 30 at the bottom of the well.

In the above-mentioned known microplates, which use small volumes of liquid, evaporation of the liquid during incubation and mixing is also particularly critical.

By using a protective film according to the invention, the microparticles can be processed in small volumes as well, since the protective film also closes the mouth of the well, thereby reducing evaporation of the liquid. Therefore, according to the invention, a separate aluminum, rubber or adhesive tape sealing cap is no longer required in microplates for mixing and incubation.

Especially when separate protective films are used in the transfer devices, the protective film may be shaped in a special way at its tip. The tip design may be designed to provide a reliable transfer of the maximum number of microparticles, for example from a viscous biological sample to another vessel. When collecting large quantities of microparticles at the tip of an elongated protective film, as occurs with a longitudinally magnetized permanent magnet, the outer 10 microparticle layers are at all times in danger of being removed and remaining in solution. Also, the liquid tension at the solution-air interface is very strong and produces a similar microparticulate release effect.

Thus, the shielding film may be shaped such that the microparticles remain as securely adhered to the shielding film as possible despite the flows generated during the movement of the transfer device, and despite the liquid surface penetration and the surface tension of the liquid surface. For this purpose, various depressions and protrusions can be made at the tip of the protective film to provide a reliable transfer of the collected microparticles to another solution. In this case, the protective film may be of either stretchable or non-stretchable material.

20 ·. The protective film of stretchable material may have a special design which ensures both reliable collection and transfer of a large number of microparticles from one container to another. For this purpose, there may be special protrusions on the edges of the protective film and; recesses into which microparticles accumulate. In this case, it is advantageous to use »· 25 transverse magnetized magnets to collect the microparticles on a large surface. The design of the protective film provides special microparticulate masses: supported structures. The design also affects the disturbing effects of fluid flows and fluid tension. When using stretchable material and different thickness points, the protrusions and recesses of the protective film are stretched in different ways. This phenomenon can be effectively utilized both for microparticle removal and especially for efficient mixing. . solution.

• · »» * 1 * 1 *, 'Concentrating large microparticle masses into smaller volumes requires efficient mixing to efficiently release the microparticles.'! 35 from the wall of the protective film. As shown, the protective film itself acts as a mixing element and is thus a highly effective device for performing mixing. Most preferably, the protective film has a different design at different locations on the protective film.

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If it is desired to collect the microparticles from the solution, the magnet is moved down and at the same time the film is stretched. When the protective film is stretched, the special design of its surface causes the accumulation of microparticles in the protected or supporting regions on the surface of the protective film. When it is desired to release the microparticles from the protective film, the magnet is moved upwardly inside the ferromagnetic sleeve. To ensure the release of the microparticles, the ferromagnetic sleeve can be moved downward, which stretches the protective film, and then upwards, and repeats these movements appropriately.

At the same time, the liquid in the vessel has been stirred very efficiently because the suitable design of the surface of the protective film 10 acts like an underwater bellows pump. Alternatively, it is also possible to move the magnet downward and thereby stretch the protective film to achieve an effective mixing based on the phenomenon described above. Moving the magnet instead of the ferromagnetic tube also causes the microparticles to move towards the magnet and the surface of the protective film, which further enhances the mixing. These methods of mixing the liquid mentioned above may also be conveniently combined. Such a mixing method also works when using magnets magnetized in the longitudinal direction.

REACTION UNIT OF THE INVENTION

The invention also relates to a microparticle reaction unit. According to a preferred embodiment of the invention, the transfer device according to the invention may also form a Reactor unit in which the vessel or reactor may be made of different materials and of variable shape. The vessel constituting the reactor chamber may have one or more openings for inlet and outlet of liquids.

•; The container may have an arrangement for recycling the liquid to be recycled for processing into a »· i» · 25 vessel. The container may be part of a larger assembly of several containers of different sizes suitably connected to each other.

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The ferromagnetic tube described in the invention may be a single tube, a plurality of more Y '; tubes together, or ice arrangements in which the individual tubes form a special formation: '*: 30 tubes. In one embodiment of the invention, the ferromagnetic tube may be special

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,:, ferromagnetic plate with one or more holes in which one or more magnets »'' can move. Such an arrangement is particularly advantageous when handling small volumes of, for example, 8, 24, 48, 96 and 384 well plate formats such as microplates or Y: s.

* ♦

Especially when dealing with very large volumes, it may be advantageous to include several magnetic units as a group of magnetic units, whereby the collection surface for large microparticle masses can be further increased. In addition, the design of the protective film can provide advantageous alternatives for handling large particle masses.

The device shown can be used to collect microparticles from a plurality of vessels or to make an arrangement whereby the fluid flows in a steady stream past the rods. The latter case has the advantage that it is relatively easy to operate even large volumes.

In both cases, the starting assumption is that the particles are first freely in solution, whereupon they are collected by the method described in the invention.

According to the invention, a plurality of magnetic rods may also be contained within a single protective film, suitably arranged on the inner periphery of the protective film. This is particularly true in the case of a very large protective film in which very large volumes of liquid are handled. Another option is to use one very large magnetic rod inside the large protective film.

15

According to the invention there may also be a solution having separately separate magnetic rods for collecting microparticles and a special device or rod for moving the liquid surface in a suitable manner as described in the invention. This solution allows for solutions in which the magnetic rods do not move at all, and the movement of the fluid and microparticles is accomplished by means of 20 specially designed members. The container used for such a solution, or. the reactor is suitably designed to meet the described needs.

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* ·; In one embodiment of the invention there are a plurality of separate magnetic rods, '; each with its own protective film. These magnetic rods may be grouped in> λ »·» 25 into a suitable formation, such as a fan in a row, a circle arc, or a plurality of nested circle arcs in which each rod collects an appropriate number of microspheres.

Ι '·' ι If such a solution is still housed in a sealed vessel or reactor which can: '': 30 add liquid as needed, and may have a separate valve from which the treated liquid '*',:, can be discharged, then the solution thus obtained can handle very large ones

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'' 'Fluid volumes. If the reactor type thus described is kept on its side, and the magnetic rod system can be rotated relative to the reactor shells, then: A: This solution can also provide a mixture for handling liquid samples and: '* 35 microparticles. The microparticles may also be pre-bonded to the magnetic rods, or may be suitably attached to the magnetic rod shield during the process so that the reactor has a very high active surface. By mixing, the liquid to be treated 115294 16 passes through the microparticles such that the desired components, such as proteins, adhere to the microparticles on the rods. On the other hand, the liquid can be entrained between the microparticles by arranging the liquid streams conveniently in the vessel or inside the reactor.

5

The device and method of the invention are not limited to, for example, molecular biology or protein purification, but are generally applicable in the fields where microparticle-bound ligands can be used to synthesize, bind, isolate, purify or enrich desired agents from various samples: diagnostic applications, biomedicine, synthesis of chemicals, isolation of bacteria and cells.

APPLICATIONS OF THE INVENTION

The device and method of the invention are suitable for use in a wide variety of applications, such as protein chemistry, molecular biology, cell biology, and proteomics. The invention has applications in industry, diagnostics, analytics and research.

In the purification of proteins, there is a need to carry out purification experiments in small volumes and, on the other hand, to increase capacity to very large volumes. With the invention described, protein purifications from different sample space volumes can be performed as needed.

Protein chemists need to be able to purify protein as little as possible from pre-treated samples such as Cell Lysate. Important t 1 ·:: It is also possible to vary the cleaning capacity according to changing needs. Today it is possible to change the column sizes used. As the purification proceeds, the protein '. · Concentration is one of the key measures. In practice, this means reducing the volume of the liquid without significant loss or denaturation of the proteins. The most commonly used methods today are dialysis or filtration, iv: Both are very time consuming methods. As described in this invention; . The device and method can provide a versatile method for varying sample volumes in the protein region. Changing capacity is easy without having to buy or make new columns. Simply select a larger number of microparticles for a larger sample volume and after binding of the proteins: Y: Remove microparticles and protein from the solution using the apparatus and method described in the invention. The washing steps can be performed either in the same container or by changing the container.

In the former case, the used wash buffers should be removed from the container and replaced with a new wash buffer. Buffer replacement can also be accomplished with various valve arrangements or 115294 17 suction arrangements. After washing, the proteins bound to the microparticles can be released in a small volume if desired and the protein solution concentrated efficiently.

Depending on the need, the volume reduction can be done in stages for a smaller volume.

5

For example, ion exchange chromatography, reverse phase chromatography, hydrophobic chromatography and affinity chromatography purifications can be performed with the device and method described in the invention. Even the apparatus described for gel filtration is capable, but requires performing actual gel filtration on, for example, 10 columns and then collecting the microparticles with the apparatus of the invention and expelling the proteins to a small volume. For example, the method allows desalting of samples without major sample dilution compared to conventional gel filtration columns.

The use of immobilized enzymes in the processing of various proteins, blind, fats and various so-called biopolymers is a very important field of application for the invention described. An important feature compared to the use of soluble enzymes is the ease of reuse of immobilized enzymes. Washing of the immobilized enzyme for further use with the described invention is very easy and efficient.

20

Examples of key enzyme groups, including those used in industry. 'Of the individual enzymes are: | - CARBOHYDRASES: Alpha-Amylases, Beta-Amylase, Cellulase, Dextranase, Alpha-Glucosidase, Alpha-Galactosidase, Glucoamylase, Hemicellulase, Pentosanase, · Xylanase, Invertase, Lactase, Pectinase, · Pullulanase. , Alkaline Protease, Bromelain, Ficin, Neutral Proteases,

Papain, Pepsin, Peptidases, Rennin, Chymosin, Subtilisin, Thermolysin, Trypsin - LIPASES AND ESTERASES: Triglyceridases, Phospholipases, Esterases, I *; '"; 30 Acetylcholinesterase, Phosphatases, Phytase, Amidases, Aminoacylase,;;, Glutaminase, Lysozyme, Penicillin Acytase · ·' - ISOMERASES: Glucose Isomerase, Epimerases, Racemases. Glucose: Y: Oxidase, Hydroxysteroid Dehydrogenase, Alcohol Dehydrogenase, Aldehyde '· · 35 Dehydrogenase, Peroxidases I »LYASES: Acetolactate Decarboxylase, Aspartic Beta-Decarboxylase, Fumarase, Histidase, DOPA 1Farboxylase 18A, Carboxylase methyltransferase,

Transaminase, Kinases - LIGASES

PHOSPHATASE: Alkaline Phosphatase 5

The use of enzymes is very common in many industries, including but not limited to the synthesis and modification of lipids, proteins, peptides, steroids, sugars, amino acids, drugs, plastics, perfumes, chemicals and so-called chiral chemicals.

10

Various synthesizing and cleaving enzymes related to glycobiology, such as endo- and exoglycosidases, are also within the scope of the invention. Similarly, enzymes known from applications in molecular biology, such as restriction enzymes, nucleases, ribozymes, polymerases, ligases, reverse transcriptases, kinases and phosphatases, are within the scope of the method described in the invention. Examples of DNA / RNA modifying enzymes include: CIAP (Calf Intestinal Alkaline Phosphatase), E. , RNase T2), DNA ligases, RNA ligases, DNA polymerases, Klenow enzyme, RNA polymerases, DNA kinases, RNA kinases, terminal transferases, AMV reverse transcriptases and phosphodiesterases. The use of these and other DNA / RNA modifying enzymes is extremely diverse as well. 'In molecular biology research and applications. In Proteomics and Protein Chemistry; ; 'Proteases are very important enzymes, some of which are trypsin; · Chymotrypsin, papain, pepsin, collagenase, dipeptidyl-peptidase IV and various' * endoproteinases. Synthetic enzymes, catalytic antibodies and V multienzyme complexes are possible for use in the methods described in the invention.

The use of the invention is also not limited by the use of enzymes and other catalytic components under anhydrous conditions, for example in organic solvents.

: 30 As concrete examples of applications of the invention in the field of molecular biology may be. . mention: *> ': / CLONING OF DNA INSERTS:: restriction enzymes are required for the cloning of DNA inserts, (eg EcoR I, Hind III, Bam. · * 35 HI, Pst I, Sal I, Bgl II, Κρη I, Xba I, Sac I, Xho I, Hae III, Pvu II, Not I, Sst I, Bgl I), creating * »blunt ends (e.g., thermostable polymerases, Klenow Fragment DNA Polymerase I, Mung Bean nuclease), ligations (e.g. T4 DNA Ligase , E. coli DNA Ligase, T4 RNA Ligase), 115294 19 phosphorylation (e.g. T4 Polynucleotide Kinase), dephosphorylation (e.g. CIAP, E. coli Alkaline Phosphatase, T4 Polynucleotide Kinase) and deletions (e.g. T4 DNA Polymerase, thermostable polymers) , Exo III Nuclease, Mung Bean Nuclease) SYNTHESIS AND CLONING OF 5 cDNA:

Reverse Transcriptase, RNase H, DNA polymerase I, T4 DNA polymerase I, E. coli DNA Ligase.

NUCLEIC ACID LABELING: 10 5 'labeling (e.g. T4 Polynucleotide Kinase), 3' addition (e.g. T4 RNA Ligase), 3 'fill-in (e.g. Klenow Fragment DNA Polymerase I, T4 DNA Polymerase), 3' exchange (e.g. .T4 DNA Polymerase, thermostable polymerases), nick translation (e.g., E. coli DNA Polymerase I, thermostable polymerases), replacement synthesis (e.g., T4 DNA Polymerase, thermostable polymerases, Exo III Nuclease), random priming (e.g., Klenow 15). Fragment DNA Polymerase I, thermostable polymerases) and RNA probes (e.g. T7 RNA Polymerase, SP6 RNA Polymerase).

NUCLEIC ACID SEQUENCE: DNA sequencing (e.g., E. coli DNA Polymerase I, Klenow Fragment DNA Polymerase I, thermostable polymerases) and RNA sequencing (eg, Reverse Transcriptase, thermostable reverse transcriptase).

MUTAGENATION OF NUCLEIC ACIDS:; ; Oligonucleotide directed (e.g. T4 DNA Polymerase, T7 DNA Polymerase, thermostable * · · * · 25 polymerases) and Misincorporation (e.g. Exo III Nuclease, Klenow Fragment DNA: '; Polymerase I, thermostable polymerases).

MAPPING:

Restriction (e.g. Exo III Nuclease), Footprinting (e.g. Exo III Nuclease); 30 and Transcript (e.g., Reverse Transcriptase, Mung Bean Nuclease).

• »» CLEANING NUCLEIC ACIDS: * »♦ * *

Isolation and purification of genomic DNA, PCR fragments, DNA / RNA probes and plasmid DNA: Y.

: \: 35 DNA DIAGNOSTIC TECHNIQUES: 115294 20 DNA Mapping, DNA Sequencing, SNP (Single Nucleotide Polymorphism), Chromosome Analysis, DNA Libraries, PCR (Polymerase Chain Reaction), Inverse PCR, LCR (Ligase Chain Reaction), NASBA (Nucleic Acid Strand-Based Amplification), Q Beta Replicase, Ribonudease Protection Assay.

5 DNA DIAGNOSTICS: RFLP (Restriction Fragment Length Polymorphism), AFLP (Amplified Fragment Polymorphism), Diagnosis of Bacterial Infections, Bacterial Antibiotic Resistance DNA Fingerprints, SAGE (Serial Analysis of Gene Expression) and DNA Sequencing.

10

The method described for isolating cells can also be used extensively. Cells of interest include, for example, stem cells, B-lymphocytes, T-lymphocytes, endothelial cells, granulocytes, Langerhans cells, leukocytes, monocytes, macrophages, myeloid cells, NK cells, reticulocytes, reticulocytes. and hybridoma cells. Well known methods such as direct or reverse cell isolation may be used for cell isolation. In the first, direct isolation mode, the desired cells are harvested separately from the sample by binding to the surface with microparticles, for example, by utilizing specific antibodies. In the indirect method, the desired cells are not bound to 20 microparticles but all other cells in the sample. In this case, the desired cells remain in solution.

«> J · * ·

Bacteria, viruses, yeasts and many other single or multicellular organisms *; method for isolation, purification and / or enrichment *! * · # 25 works well. A particularly important field of application is for pathogenic bacteria such as e.g.

> · Enrichment of large volumes of salmonella, Listeria, Campylobacter, E. coli 0157 and Clostridium, viruses, parasites, protozoa or other small organisms. The device and method described in the invention can also be utilized in these application areas.

1 ·: '': 30 Biocatalysis is commonly understood to mean enzymes of bacteria, enzymes or other

> * I

containing components in the process. Enzymes or bacteria may be immobilized on a suitable solid support and the material to be treated is contacted with the immobilized components, for example, using conventional columns. According to this invention, the cells or enzymes may conveniently be attached to microparticles, which are then used in accordance with the invention to perform various enzymatic reactions.

115294 21

Isolation of cellular organelles and various cellular fractions is also within the scope of the invention. Cellular organelles can be isolated in the normal manner by utilizing, for example, specific antibodies or various affinity ligands.

5 Purification of nucleic acids has a wide variety of needs, ranging from the purification of very small amounts of DNA (Deoxyribonucleic Acid), RNA (Ribonucleic Acid) or mRNA (Messenger RNA) to the high processing requirements of many liters. The method of the present invention can efficiently isolate nucleic acid from both large and small sample volumes.

10

The method can be used to concatenate isolation and cleaning processes according to different needs. For example, one may first isolate the desired cells from the sample and purify them.

Thereafter, for example, cellular organelles can be isolated from the cells. The cellular organelles are purified and the process can continue, for example, to purify DNA or protein.

15 During the process, microparticles with different coatings and properties can be varied according to the needs. The final step is to concentrate the purified product to the desired volume.

BRIEF DESCRIPTION OF THE DRAWINGS

Figures 1A-1G schematically illustrate a microparticle transfer device · according to the invention. different embodiments of the magnetic unit cut.

Figures 2A-2G schematically illustrate various embodiments of magnets in a magnetic unit and their magnetic fields.

Figures 3A and 3B schematically illustrate embodiments of a magnetic unit placed in a solution containing microparticles.

Figures 4A-4B correspond to Figures 3A and 3B and show other ... embodiments of a magnetic unit in solution.

Figures 5A-5E illustrate embodiments of a magnetic unit with a non-stretchable protective film and a longitudinally magnetized magnet in solution.

Figures 6A-6E illustrate embodiments of a magnetic unit with a non-stretchable protective film and a transverse magnetized magnet in solution. Figures 7A-7E show a stretchable protective film and longitudinally magnetized. embodiments of a magnetic unit with a magnet in solution.

Figures 8A-8E show embodiments of a magnetic unit having a stretchable protective film and a transverse magnetized: 35 magnet in solution.

Figures 9A-9G show the steps of using a magnetic unit to transfer microparticles from one vessel to another.

115294 22

Figure 10 is a side view and a sectional view of a hand-operated microparticle transfer device.

Figure 11 is a side elevational view and a sectional view of a manually operated microparticle multi-channel transfer device.

Figure 12 schematically shows a transfer device automaton.

Figure 13 shows a further embodiment of the magnetic unit, seen from the side and partially cut away.

Figure 1.4 is a side elevational view and a sectional view of a reactor vessel according to the invention.

Figure 15 is a side elevational view and partially sectional view of a reactor unit according to the invention.

Figure 16 shows the reactor unit of Figure 15 in horizontal position.

Fig. 17 shows a perspective view of an environmental cabinet according to the invention.

Fig. 18 is a side view and sectional view of a tube.

Fig. 19 is a side elevational view and sectional view of a magnetic unit in connection with a test tube having a sleeve in a first position.

Fig. 20 corresponds to Fig. 19 and shows a situation where the sleeve of the magnetic unit is in a second position.

Fig. 21 corresponds to Fig. 19 and shows a situation in which the sleeve 20 of the magnetic unit is in the third position.

Fig. 22 corresponds to Fig. 18 and shows the test tube in another situation, f Fig. 23 shows a side view and partially cut away of a magnetic unit '(*: embodiment) with a different protective film.

; '; Fig. 24 corresponds to Fig. 23 and illustrates the operation of the magnetic unit in a second step.

Fig. 25 corresponds to Fig. 23 and shows the operation of the magnetic unit in the third step.

> ·

Figure 26 corresponds to Figure 23 and shows the operation of the magnetic unit in the fourth step.

Fig. 27 shows yet another magnetic unit having a protective film; * 30 embodiments seen from the side and partially cut.

• *

Fig. 28 is a schematic side elevational view and a sectional view of a plurality of parallel: magnetic units having a common sheet-shaped protective film; Fig. 29 corresponds to Fig. 28 and shows parallel magnetic units according to another embodiment.

Fig. 30 corresponds to Fig. 28 and shows parallel magnetic units according to a third embodiment.

115294 23

Figure 31 corresponds to Figure 28 and shows parallel magnetic units according to a fourth embodiment.

Figure 32 is a schematic top view of parallel magnetic units disposed in a circular shape.

5

DETAILED DESCRIPTION OF THE DRAWINGS

Fig. 1A shows an embodiment of the magnetic unit 10 according to the invention. including a ferromagnetic tube or sleeve 12 having a rod-shaped permanent magnet 13 movable through a rod or transfer pin 11. The junction 10 between the magnet 13 and the bar 11 is denoted by reference numeral 14 and the aperture at the end of the tube 12 is denoted by the reference numeral 15. By moving the bar 11 and the inside tube 12 axially with respect to one another and the magnet 13 attached thereto may be moved within the tube 12, or the tube 12 may be moved so that the bar 11 and magnet 13 remain in place. Alternatively, both parts 12 and 13 may also move. In all these alternative ways, the magnet 13 is forced out of the opening 15 at the end of the tube 12 and again in the tube 12.

In Fig. 1A, the diameter of the rod 11 is larger than the diameter of the magnet 13. The magnet 13 20 is connected to the bar 11 so that the end of the magnet 13 is inserted into the recess at the end of the bar 11. There is a sufficiently tight fit between the cavity and the magnet 13 to hold the magnet 13 and the rod connected. Since the internal diameter of the ferromagnetic tube 12 in this I solution is larger than the diameter of the magnet 13, in some cases it may '; be harmful.

25

Fig. 1B shows another embodiment of the magnetic unit 10, wherein the diameters of the magnet 13 <and the rod 11 are equal. The connecting member between the magnet 13 and the rod 11 is a thin-walled sleeve 16 into which the ends of both the rod 11 and the magnet 13 are inserted. The inner diameter of the thin-walled sleeve 16 is formed such that the fit between the sleeve 16: '': 30 and the magnet 13 and the fit between the sleeve 16 and the bar 11 are tight enough to hold these parts together. Since the sleeve 16 is thin-walled, the diameter of the magnet 13 may be nearly as large as the inside diameter of the ferromagnetic tube 12-1.

Fig. 1C shows a third embodiment of the magnetic unit 10, wherein the mouth 15 of the end of the ferromagnetic tube 12 is reduced. In this way, the clearance between the magnet 13 115294 24 and the tube 12 is obtained, even if the inner diameter of the tube 16 is clearly larger than the diameter of the magnet 13.

Fig. 1D shows a fourth embodiment of the magnetic unit 10, wherein the connection 14 between the magnet 13 5 and the rod 11 is made by glue. In this solution, the diameters of the magnet 13 and the rod 11 are equal, so that the distance between them and the inner surface of the tube 11 can be suitably small.

Fig. 1E shows a fifth embodiment of the magnet unit 10, wherein the magnet 13 10 and the rod 11 are joined together by the magnetic force of the magnet 13 such that the magnet 13 pulls the rod 11 sufficiently tight against the magnet 13. The solution only works if the rod 11 is ferromagnetic material. Also in this solution, the diameters of the magnet 13 and the rod 11 are the same.

Fig. 1F shows a sixth embodiment of the magnetic unit 10 with a projection formed at the end of the rod 11 which is inserted into the recess formed at the end of the magnet 13. At the junction 14, the fit between the projection and the cavity is made tight enough to hold these parts together.

Figure 1G shows a seventh embodiment of a magnetic unit 10 having an electromagnet instead of a permanent magnet. In this solution, the rod 11 is of ferromagnetic material, and a coil 27 is disposed about its other end, which t induces a magnetic field on the rod 11 when the voltage source is connected to the coil 27. Thus; thus, the rod 11 acts as an electromagnet, whereby a separate permanent magnet to be attached thereto is not required.

». <: Fig. 2A shows an embodiment of the magnetic unit 10, in which the mounting end of the magnet 13 corresponds to the solution of Fig. 1B, i.e. the magnet 13 is connected to the bar 11 by a sleeve. However, no magnetization direction of the magnet was mentioned in Figure 1B.

In the magnet unit 10 of Figure 2A, the magnetization direction of the magnet 13 is parallel to the longitudinal axis of the magnet 13 't.

. The embodiment of the magnetic unit 10 shown in Figure 2B corresponds to the solution of Figure 2A in other respects, but the magnetization direction of the magnet 13 is transverse, i.e. perpendicular; 35 magnets against 13 longitudinal axes. However, in both Fig. 2A and Fig. 2B, the magnet 13 may also be attached to the rod 11 in any other manner.

115294 25

Figures 2C-2G schematically show the magnetic field provided by the magnet 13 of the magnet unit 10 in various embodiments.

Fig. 2C shows the magnet 13 of the magnet unit 10 being magnetized 5 in the longitudinal direction, as in Fig. 2A. 2C, the other end of the magnet 13 is partially pushed out of the tube 12, whereby its magnetic field 17 extends from the distal end of the magnet 13 to the end of the tube 12. The maximum magnetic flux density for this solution is obtained around the free end of the magnet 13, the area of which is indicated by reference numeral 18 in Figure 2C. The described solution causes the microparticles to accumulate substantially at this end of the magnet 13 only, thereby limiting the amount of

Fig. 2D shows the magnetic field of the magnet 13 of the magnet unit 10 when the magnetisation axis of the magnet 13 is transverse, i.e. according to Fig. 2B. In this case, the resulting magnetic field 19 is uniformly distributed over the entire magnet 13, thereby maximizing the microparticle collection surface.

However, if it is desired to delimit the collection surface of the magnet 13 of the magnet unit 10, the magnet 13 may be partially enclosed within the ferromagnetic tube 12. Such a situation 20 is shown in Figure 2E. In this case, the collecting surface 20 of the magnet 13 is less than,. 2D.

»· 1» «I ·! Figures 2F and 2G are schematic cross-sections of two different transverse '; the magnet 13 of the magnetized magnet unit 10. In Figure 2F, the magnet 13 is divided into two parts in a plane parallel to the longitudinal axis'! In Figure 2G, the magnet 13 is divided into four longitudinal portions, respectively. Figures 2F and 2G show that ...: the magnetic fields are different in them because the magnetic fields are positioned slightly differently.

However, both solutions and all their variations are equally applicable.

3A shows a magnetic unit 10 for collecting microparticles 22 in a container 26, such as a solution 23 in a test tube 23. The magnet 13 protected by the protective film 21 is attached to a bar 11 which is not ferromagnetic. In Figure 3A, the magnet 13 is completely below the liquid surface 25 so that the distance of the magnet 13 from the liquid surface 25 is h. The magnet 13 in Figure 3A is magnetized along the longitudinal axis 35 of the magnet 13. The microparticles 22 then accumulate from the solution 23 in the vessel 26 at each of the poles 24a and 24b of the magnet 13 outside the shield 21, both at the tip of the shield 21 and at the junction 14 of the rod 11 and magnet 13. This 115294 26 is normal 25 below.

Fig. 3B shows another embodiment of the magnetic unit 10, which also includes a magnet 13 protected by a shielding film 21 which is completely below the liquid surface 25 at a distance h from the liquid surface 25. This embodiment corresponds to the embodiment of Fig. 3A in other respects. Figure 3B shows that microparticles 22 now accumulate over a large area outside the protective film 21. However, it would be most advantageous to have all the microparticles 22 collected at the very bottom of the tip of the 10 magnetic units. It is particularly advantageous when the microparticles 22 are to be transferred to a small volume of liquid. In Figure 3B, the microparticles 22 do not accumulate in a small area, and in particular near the lower part of the protective film 21. Therefore, this option is not particularly advantageous when it is desired to concentrate the microparticles 22 into small volumes of liquid.

15

Figure 4A shows the magnetic unit 10 disposed in solution 23 in test tube 26 and the accumulation of microparticles 22 near the lower part of the magnets 13 protected by the protective film 21 of the magnetic unit 10. In Fig. 4A, magnet 13 and both magnetic poles 24a and 24b are completely below the liquid surface 25. However, the microparticles 20 do not accumulate anywhere other than the lower part of the protective film 21, because the magnet 13 *: the upper pole 24b has been successfully short-circuited by conveniently placing the ferromagnetic tube 12 on the magnet 13. At the upper pole 24b of the magnet 13, there is no magnetic field outside the ferromagnetic tube 12, which is why microparticles 22 are not detected outside the shielding film 21. The described magnetic unit 10 can concentrate 25 microparticles 22 into small liquid volumes. 'Below the liquid surface 25 and is fixed to a non-ferromagnetic bar 11.

* ·

i «I

When, in the situation shown in Fig. 4A, the magnet 13 is moved completely inside the ferromagnet tube 12, the magnetic field of the magnet 13 is almost completely removed.

* »♦ 30 The microparticles 22 can thus be released from the surface of the protective film 21 simply by inserting the magnet 13 completely inside the ferromagnetic tube 12. Microparticles 22 ''. can be transferred from vessels 26 to one another bound to the surface of the shielding film 21, whereby the magnet 13 is conveniently located outside the ferromagnetic tube 12.

Fig. 4B shows a magnetic unit 10 corresponding to the embodiment of Fig. 4A in other respects but with the magnet magnetized transversely. In Fig. 4B, the magnetic field 13 of the transverse magnetized magnet 13 is reduced by 115294 27 by ferromagnetic tube 12. In the situation shown in Fig. 4B, the magnet 13 is very little outside the ferromagnetic tube 12. Figure 4B shows that the transverse magnetized long magnet 13 and the shield tube 12 can simply concentrate the microparticles 22 to the very bottom of the shielding film 21. Thus, Figures 4A and 4B each show advantageous and effective methods and devices for treating microparticles in small fluid volumes.

Figures 5A-5E illustrate a stepwise collection of microparticles 22 from solution 23 by means of a magnetic unit 10 provided with a non-stretchable protective film. The magnet 13 and the ferromagnetic tube 12 are movable axially relative to one another and magnet 13 is magnetized along its longitudinal axis.

Figures 5A-5E also show various ways of concentrating microparticles by means of a ferromagnetic tube 12 and a magnet 13 at the very bottom of the protective film 21 and release them, for example, into small volumes of liquid.

Fig. 5A shows a magnetic unit 10 whose magnet 13 is ejected from the ferromagnetic tube 12 by a non-ferromagnetic rod 11, the magnetic field of the magnet 13 being substantially in the lower part of the protective film 21. In this case, the microparticles 22 20 also accumulate in the lower part of the protective film 21. Also, in the following examples, the magnet moving rod 11 is not ferromagnetic.

Fig. 5B shows the magnet unit 10 of Fig. 5A with its magnet 13! in another position. In Figure 5B, the magnet 13 is moved almost completely into the ferromagnetic tube 12 while the tube remains stationary. In this case, some of the microparticles 22 in solution 23 move upward along the protective film 21.

Fig. 5C shows the magnetic unit 10 of Fig. 5B with its magnet 13 fully retracted into the tube 12, whereby the microparticles 22 are dispersed in solution: 30 23. Thus, when the magnet 13 is moved upwardly from the lower part of the protective film 21, best possible to collect microparticles 22 on the side portion of the protective film 21. It is due to the magnet's 13 magnetic fields and its magnetic poles. position and attraction with respect to the protective film 21 used. Thus, this alternative: is possible, but not the most advantageous, for detaching microparticles from the surface of the protective film 21.

However, by optimizing the microparticles 22 and the upward motion of the magnet 13, a good result can be obtained, i.e. the microparticles remain very close to the lower part of the protective film 21 28 115294

Figure 5D shows an alternative and effective way to release microparticles 22 in a controlled manner from the lower portion of the protective film 21 of the magnetic unit 10 of Figure 5A, for example to small volumes. Instead of the upward movement of the magnet 13 shown in Figure 5B, the ferromagnetic tube 12 is now moved downwards in Figure 5D. The figure shows that the microparticles 22 do not move upwardly along the protective film 21.

Fig. 5E shows the magnetic unit 10 of Fig. 5D with the ferromagnetic tube 12 completely transferred to the magnet 13. The figure shows that the microparticles 22 thus remain better in solution 23 in the lower part of the test tube 26 near the end of the magnetic unit 10.

However, neither of the methods shown in Figures 5B-5C or 5d-5E is particularly advantageous for the collection and processing of very large microparticle masses.

15

Figures 6A-6E show a stepwise collection of microparticles 22 by means of a magnetic unit 10 provided with a non-stretchable protective film 21 in which the magnet 13 or the ferromagnetic tube 12 is moved and when the magnet 13 is transversely magnetized.

Fig. 6A shows a magnetic unit 10 with a transverse magnetized magnet 13 being ejected from a ferromagnetic tube 12 which covers only a small portion of the magnet 13. Thus, the microparticles 22 accumulate outside the protective film 21 of the magnetic unit 10.

Fig. 6B shows the magnetic unit 10 of Fig. 6A in a position where the magnet 12 is moved up almost completely inside the ferromagnetic tube 12. In this case, most of the microparticles 22 at the bottom of the protective film 21 move upward with the magnet 13.

Figure 6C shows the magnetic unit 10 of Figure 6B in a position where the magnet 13 is completely contained within the ferromagnetic tube 12. Thus, the microparticles 22 are released into the surrounding solution 23. Thus, this method is not suitable for concentrating the microparticles 22 to the lower part of the protective film 21 and transferring it, for example, to a small volume of liquid.

, ': 35 115294 29

Fig. 6D shows the magnetic unit 10 of Fig. 6A in a position in which the ferromagnetic tube 12 is moved downwardly almost completely over the magnet 13. At the same time, the microparticles 22 suitably move downwardly with the tube 12.

Figure 6E shows the magnet unit 10 of Figure 6D in a position where the magnet 13 is completely covered by the ferromagnetic tube 12. The figure shows that in this way the microparticles 22 can be effectively concentrated near the lower part of the protective film 21 of the magnetic unit 10. Thus, this solution is well suited both for collecting large volumes of microparticles and for concentrating microparticles to small volumes of liquid.

Figures 7A-7E show a stepwise collection of microparticles 22 by means of a magnetic unit 10 provided with a stretchable protective film 21 by moving either the magnet 13 or the ferromagnetic tube 12. The magnet 13 is magnetized 15 in the longitudinal direction.

Fig. 7A shows a magnetic unit 10 in which the longitudinally magnetized magnet 13 is ejected from the ferromagnetic tube 12 while simultaneously stretching the stretchable protective film 21. The microparticles 22 then accumulate near the end of the magnet 13 at the bottom of the stretched protective film 21. At the same time, due to the stretching of the protective film 21, the thickness of the protective film 21 is reduced, whereby the magnetic field is>> · increased with the thinning of the protective film 21.

! Fig. 7B shows the magnetic unit 10 of Fig. 7A in a position where the magnet 13 25 is moved upwardly inside the ferromagnetic tube 12. At the same time also stretched ,,; 'The protective film 21 returns upwards. It follows that a sufficient magnetic field is still applied to the lower portion of the upwardly movable protective film 21 to keep the microparticles 22 accumulated on the protective film 21.

Fig. 7C shows the magnetic unit 10 of Fig. 7B in the position where the magnet 13 is. completely drawn inside the tube 12 and the microparticles 22 are released from the magnetic field. In this way, the microparticles 22 can be well concentrated to the lower part of the protective film 21 and further transferred to a small volume of liquid.

Fig. 7D shows the magnetic unit 10 of Fig. 7A in a position in which the ferromagnetic tube 12 is moved downwardly over the magnet 13. The magnet 13 does not move but keeps the protective film 21 further stretched. Due to the stretching of the protective film 115294 30, the magnetic field is very large and the microparticles 22 remain very well adhered to the protective film 21.

Fig. 7E shows the magnet unit 10 of Fig. 7D in a position where the ferromagnetic tube 12 is completely moved onto the magnet 13. This removes the magnetic field and releases the microparticles 22 into the liquid 23. This method is very well suited for concentration in small liquid volumes.

Figures 8A-8E illustrate a stepwise collection of microparticles 22 by means of a magnetic unit 10 provided with a stretchable protective film 21 by moving either a magnet 13 or a ferromagnetic tube 12. The magnet 13 is transversely magnetized.

Fig. 8A shows a magnetic unit 10 in which the transversely magnetized magnet 13 is ejected from the ferromagnetic tube 12 so that it at the same time stretches the stretchable protective film 21. The microparticles 22 then accumulate over a very large area around the stretched protective film 21 by the magnet 13.

Fig. 8B is a view showing the magnetic unit 10 of Fig. 8A in a position where the magnet 13 is moved upwardly inside the ferromagnetic tube 12. When the magnet 20 13 is moved upwards, the stretched protective film 21 will at the same time return to its original shape, i.e. with the magnet 13 upwards. Microparticles 22 move with and across * ·. the microparticle mass may be concentrated in a small area at the tip portion of the protective film 21.

! Fig. 8C is a view showing the magnetic unit 10 of Fig. 8B in a position where the magnet 13 is completely retracted inside the ferromagnetic tube 12. the microparticles 22 are released from the magnetic field into the solution 23.

Fig. 8D shows the magnetic unit 10 of Fig. 8A in a position in which the ferromagnetic tube 12 is moved downwardly over the magnet 13. Here, as in Figures 8B and 8C, the microparticles 22 •: 30 can be collected from a large sample volume and concentrated • in a small area at the tip of the protective film.

Fig. 8E is a view showing the magnetic unit 10 of Fig. 8D in a position where the ferromagnetic tube 12 is moved over the magnet 13 completely. In this case, the 1: 35 magnetic field is removed and the microparticles 22 are released from the magnetic field into solution 23.

115294 31

Figures 9A-9G illustrate a stepwise application of a magnetic unit 10 for collecting a large microparticle mass from a high volume of liquid and concentrating the microparticles into a small volume.

Figure 9A shows a container 26a in which the microparticles 22 are contained in the liquid 23 in a large volume.

Fig. 9B shows a magnetic unit 10 according to the invention, which is disposed in the container 26 of Fig. 9A. The magnetic unit 10 of Fig. 9B has a magnet 13 protected by an unstretchable protective film 21 which is magnetized transversely. With such a magnetic unit 10, the microparticles 22 are collected in a large area on the surface of the protective film 21.

Figure 9C shows another container 26b having a small volume of liquid 23b. To this vessel 26b, microparticles 22 collected from the vessel 26a of FIG. 9A by the magnetic unit 10 are transferred to the vessel 26b of FIG. 9C, suitably selected for use with the magnetic unit 10 shown.

Figures 9D-9F illustrate a step-by-step process of releasing microparticles 22 from a large volume to a small volume.

9 · Fig. 9D shows the magnetic unit 10 embedded in the vessel 26b. the object of immersing the magnetic unit 10 in liquid 23b. , The liquid surface of the liquid volume is suitably raised beyond its upper limit; 'Microparticles 22 are collected from the large vessel 26a shown in Fig. 9B.

'· · ·' The method utilizes the fact that, when immersed in a liquid, the object displaces one volume of liquid. When a suitable mile and shape receptacle is used, as well as a magnetic unit suitable for its size and shape, the liquid surface in the receptacle: 30 rises to the desired height. What is important here is that the particles remain below the surface of the liquid.

Fig. 9E shows the magnetic unit 10 of Fig. 9D in a situation where: the ferromagnetic tube 12 is moved downwards. The microparticles 22 are then released, '35 downwards from the surface of the protective film 21.

115294 32

Fig. 9F shows the magnetic unit 10 of Fig. 9E in the following situation, in which the ferromagnetic tube 12 is moved completely over the magnet 13 and there is no longer a magnetic field outside the tube 12 to hold the microparticles 22 on the surface of the protective film 21. The microparticles 22 are now completely released into surrounding fluid 5b.

Fig. 9G shows a situation in which the magnetic unit 10 has been moved out of the vessel 26b, whereby the liquid surface has lowered back to the initial state. As a result, the high microparticulate mass has been efficiently and simply transferred to a small volume of 10, as shown in Figure 9G. From this, concentration can be continued as described above or using the methods shown in the previous figures. The transfer and concentration steps of the microparticles 22 may be suitably carried out in various ways as required.

Fig. 10 shows an example of a manually operated microparticle transfer device 30 according to the invention. The transfer device 30 comprises a body tube 31, an extending adapter sleeve 32 and a magnetic unit 10 according to the invention at the end of the transfer device. The magnetic unit 10 has a magnet 13, a rod or transfer pin 11, a ferromagnetic tube 12 and a stretchable or rigid protective film 21 printed on the adapter sleeve 32.

The non-ferromagnetic rod 11 which moves the magnet 13 of the magnet unit 10 extends up to the top of the transfer device 30 where it is connected to the magnet transfer bar 37. This transfer bar 37 is manually moved by a magnet transfer pin 38 which protrudes out of the wall 31 of the body tube 31. The magnet transfer pin 38 can be pushed up, down and down in the opening 39, whereby the transfer slide 37 and the rod 11 and. . The magnet 13 moves up and down.

Further, the microparticle transfer device 30 also has a mechanism for moving the ferromagnetic tube 12 in the axial direction. The mechanism includes a tube transfer unit 34 and., A tube transfer pin 35, which also protrudes through the body tube 31 from another elongated * ·,: and down in opening 36, * ·. wherein the tube transfer unit 34 and also the ferromagnetic tube 12 move up and down.

The microparticle transfer device 30 is held in a hand so that the finger can easily: 35 move both the magnet transfer pin 38 and the tube transfer pin 35.

115294 33

Fig. 11 illustrates an example of a hand-operated microparticle multi-channel transfer device 40 having a magnetic unit group 41 comprising eight magnetic units 10 according to the invention. Magnetic units 10 of a magnetic unit group 41 are in a row. Each magnet unit 10 has a magnet 13, a transfer pin 11, 5, a ferromagnetic tube 12 and a protective film 21. In the example shown in Fig. 11, no mechanism is shown for moving the ferromagnetic tubes 12 of the magnetic units 10 up and down as in the previous example. By way of example, the figure shows only a simple mechanism for simultaneously moving all eight magnets 13 of the magnetic units 10.

10

In Fig. 11, the mechanism of the magnets 13 of the magnet units 10 includes a connecting rod 43 into which the rods 11 of all the magnets 13. is attached. The magnets 13 of the multichannel transfer device 40 are moved downwardly and out of the ferromagnetic tubes 12 by pressing with a finger a "trigger" 46 outside the transfer device which is connected to the connecting rod 43 of the magnets 13 via the intermediate rod 45 15.

According to one embodiment of the microparticle multi-channel transfer device 40, all the magnets are simultaneously, but that, if necessary, some of the magnets 13 can be locked in the desired position. In addition, the various magnetic units 10 may have a mechanism by which the ferromagnetic tubes can also be moved up and down.

• •. Fig. 12 shows a microparticle transfer device 50 having magnetic units according to the invention in a row or n x m matrix 25 51 shown in Fig. 12. Magnetic units 10 are attached to a control unit 52 having the necessary mechanics f for moving magnets and ferromagnetic tubes vertically. The control unit 52 may also move upwards and downwards in the direction of the arrow 54 and / or the direction of arrow 53 according to the lateral direction. Below the magnetic units, a sample plate 55 is placed over the plane 57 either manually or by means of a laboratory robot. The sample plate 55 has sample wells either in one row: 30 in a row or in a matrix 56 as shown in Figure 12. The automaton 50 further comprises a second control unit 58 which manages the transfer logic and contains all the necessary electronics to control the actuators and interact with other laboratory equipment.

| Fig. 13 shows a magnetic unit 10 according to the invention having a transversely magnetized magnet 13 and a ferro-metal tube or sleeve 12 which is axially displaceable on the magnet 13. The magnet 13 is protected by a protective film 21, which may be 115294 34 of a stretchable or hard material, most preferably plastic or silicone rubber. In addition, the magnet unit 10 includes a latching flange 33 and a rotation axis 28, by means of which the magnet 13 and the protective film 21 inside the magnet unit 10 can be rotated about their longitudinal axes.

5

Figure 14 illustrates a reactor vessel 61 according to the invention having passageways 62 provided with valves 63. Reactor vessel 61 has a fluid 23 in the process. Reactor vessel 61 and magnetic unit 10 shown in Figure 13 together form reactor unit 60 as shown in Figure 15.

10

Figure 15 shows a reactor unit 60 according to the invention, in which a reactor vessel 61 is provided with the required solution 23 in the process, such as culture medium, sample, buffer solution and magnetic particles 22 such as microparticles. Thereafter, the reactor vessel 61 is connected to the mounting flange 33 of the magnetic unit 10. Further substances, such as suitable solutions and magnetic particles, may be added to the reactor 60 as required, or liquids removed through channels 62 with valves 63 connected to the reactor vessel.

The passages 62 or the corresponding inlets may be located at the sides or ends of the reaction vessel and may be provided in multiple locations throughout the reactor unit. Channels 62 can be used to control, for example, gases, pH and salinity within the reaction unit 60.

Through the inlet channels 62, additional sample can also be introduced into the reaction unit 60 and / or: the sample inside the reaction unit 60 can be removed. These inlets may have suitable filters, whereby the gas or solution to be introduced can also be kept sterile. In Fig. 15, magnetic particles 22 have accumulated on the surface of the protective film 21.

Fig. 16 is a horizontal view of the reactor unit 60 of Fig. 15. If the reactor unit 60 is kept on its side in this position and the magnet 13 of the magnet unit 10 and the shielding film 21 are rotated relative to the sheath of the magnetic unit 10, an effective mixing of the fluid 23 inside the reactor unit 60 is achieved. The magnetic particles are also mixed with the liquid. The height of the liquid surface 25 inside the reactor unit 60 30 can be adjusted and optimized according to the application used.

In order to enhance the mixing of the liquid 23 in the reactor unit 60, the protective film 21 of the magnet 13 may be provided with suitable flaps. As the shielding film 21 and the blades 35 rotate, the liquid 23 in the reactor vessel 61 is effectively agitated and agitated. Instead of the flaps, the surface of the protective film 21 may also be shaped in various ways. The shielding film 21 may also have a suitable design at its tip portion 64, which then supports the magnetic unit when positioned horizontally on its side.

In the process used, the magnetic particles may be pre-bonded to the protective film 21 of the magnet 13 5 or may be conveniently attached thereto during the process.

The collection and removal of the magnetic particles from the protective film 21 is carried out according to the invention by means of a ferromagnetic sleeve 12 which is longitudinally moved on or off the magnet 13. The magnet 13 used in the embodiment shown is a transverse magnetized magnet. Here, it is essential that the magnetic particles 10 can be collected in the reaction unit 60 on a large surface around the protective film 21.

When magnetic particles are adhered to protective film 21, the device has a very active surface for collecting proteins, cells, DNA or bacteria from solution 23 in reaction vessel 15 61. By stirring the solution, the solution to be treated passes through magnetic articles adhered to protective film 21. It is also possible for the reaction unit 60 to occasionally release the magnetic particles into the solution as described in the invention and pick up the magnetic particles again from the solution on the surface of the protective film 21.

20

Fig. 17 shows a condition cabinet 70 according to the invention in which a plurality of · ·; : 60 reaction units simultaneously. By means of the motor 71 and the actuator 72 connected to the condition cabinet 70, several reaction units 60 can be simultaneously operated; : ': the magnets 13 and the protective films 21 inside the condition cabinet 70 are adjustable; 1 · · 25, including its temperature, rotation speed of magnets and their protective films,. exchange of gases inside the reactor units, sampling of the reactor units and sampling,. or addition of solutions to reactor units.

»· * · I, Such a solution is particularly useful in microbiological quality control, where t>» ;; For example, pathogenic bacteria can be grown in reaction units 60. After a suitable time; 1 ', the reactor units 60 are removed from the ambient cabinet 70. Then, the magnetic particles: are contained in the magnetic unit 10, collected on the surface of the protective film 21. The magnetic unit 10 of the reactor 60 is removed from the reaction vessel 61, after which the magnetic particles can, for example, be washed and concentrated in separate vessels. In the detached reaction vessel 61 35 all but the magnetic particles remain. The equipment can handle very large volumes of liquid.

115294 36

Figure 18 shows a tube 26 with a suitable liquid 23, such as a washing liquid. The magnetic unit 10 removed from the reactor 60 is introduced into the test tube 26 as shown in Figure 19. The magnetic particles 22 are then still deposited on the surface of the protective film 21. In this situation, the liquid surface 25 of solution 23 must be above the binding region of the magnetic particles 22 on the surface of the protective film 215 so that the magnetic particles 22 are below the liquid surface 25.

Fig. 20 shows a situation in which the ferromagnetic sleeve 12 of the magnetic unit 10 is moved downwards in the figure. Figure 20 shows that the ferromagnetic sleeve 12 is already partially over the magnet 13. Moving the ferromagnetic sleeve 12 over the magnet 13 causes the magnetic field to be removed at the point whereby some of the magnetic particles 22a are released from the surface of the protective film 21 from above. At the point where the ferromagnetic sleeve 12 is not yet on the magnet 13, a second portion of the magnetic particles 22b is still held by the magnetic field on the surface of the protective film 21.

15

In Fig. 21, the ferromagnetic sleeve 12 has already moved completely over the magnet 13. In this case, the ferromagnetic sleeve 12 has caused the magnetic field to disappear completely, whereby all the magnetic particles 22 have been released from the surface of the protective film 21 into the solution 23.

20: In Figure 22, magnetic unit 10 has been removed from test tube 26, whereby magnetic particles 22 and associated components such as bacteria have been concentrated in test tube 26 outside reaction unit 60. Using the same magnetic unit 10, processing of the sample in smaller volumes a bonding region of the magnetic particles 22 directly to the tip portion of the protective film 21. From test tube 26, magnetic particles 22 'can be collected in subsequent test tubes and, for example, washed as required. It is also possible to isolate the DNA, RNA, protein 30 or surface antigen of the bacteria collected from the reaction unit 60 with the reagents specifically intended for them. Bacteria must generally be: disintegrated by various devices and / or reagents prior to further analysis. After digestion, the following magnetic particles of different binding properties can be added to the bacterial lysate obtained above. For example, magnetic particles containing the novel property are used to collect the desired bacterial protein, antigen, DNA, rRNA, RNA or mRNA from bacterial lysate. In the reactor unit 60, the magnetic particles 22 for collecting bacteria may have been removed before the magnetic particles having novel properties have been introduced into the process.

115294 37

Using the method described in the invention, the above components may be isolated, washed and released for actual analysis. The assay methods may be, for example, PCR amplification or ELISA assay. In a reactor vessel as described, both aerobic and anaerobic microorganisms can be grown.

Fig. 23 shows a magnetic unit 10 comprising a transverse magnetized magnet 13, a ferromagnetic sleeve 12 and a protective film 21 having ridges 29 on the outer surface thereof, depressions between which the microparticles 22 10 accumulate and ensure reliable collection of a large number of microparticles on a large surface to move them from one container to another.

Fig. 24 shows the magnetic unit 10 of Fig. 23 in a position where the magnet 13 is completely pushed out of the ferromagnetic sleeve 12. In this case, the transversely magnetized magnet 13 collects microparticles 22 on the protective film 21 over the entire length of the magnet. As the magnet 13 is ejected, the protective film 21 is at the same time stretched so that large recesses or pockets are formed between the ridges 29. The microparticles 22 remain in these pockets so that they can be easily held in place when the magnetic unit 10 is lifted. The fluid flows caused by the motion of the magnetic unit 10 and the disruptive effect of the fluid tension 20 caused by the permeation of the surface do not release the microparticles from the pockets 22.

Fig. 25 illustrates a situation in which the magnet 13 is completely protruded from the ferromagnetic sleeve 12 and the ferromagnetic sleeve 12 is also pushed>; completely out. In this case, the ferromagnetic sleeve 12 inserted on the magnet 13 cancels the magnetic force of the magnet 13 and the microparticles 22 release from the protective film and transfer back to the liquid.

Fig. 26 again shows a situation in which only the ferromagnetic sleeve 12 is fully extended. Again, the magnet 13 has no magnetic force, so that microparticles 22: 30 do not accumulate on the surface of the protective film 21. Instead, this step of Fig. 26 can be used alternately with that of Fig. 23 to provide an effective mixing pumping effect in the liquid. Of course, the phases of Figures 24 and 25 can also be used alternately, i.e., with the magnet 13 fully extended, only the ferromagnetic sleeve 12 is moved back and forth.

.: 35 This also gives the mixing pumping effect in the liquid.

38 115294

Fig. 27 shows a magnetic unit 10 having a longitudinally magnetized magnet 13, a ferromagnetic sleeve 12 and a protective film 21 with a pocket 42 at its end for microparticles 22. Such a structure also allows a large number of microparticles to be collected, which do not readily detach from the surface of the protective film 21 during transfer.

5

Figure 28 illustrates a plurality of parallel magnetic units 10 having a common sheet-shaped protective film 21. The protective film 21 is a stretchable material, whereby the same film may be used jointly for adjacent magnetic units 10. The film is preferably rolled, so that it is also easily replaceable.

10

Figure 29 shows two parallel magnetic units 10a and 10b having a common protective film 21. In the exemplary apparatus of Figure 29, the operation of the magnetic units 10a and 10b is in different stages. The ferro-metal sleeves 12a and 12b of both magnetic units 10a and 10b are pressed against the protective film 21 such that the protective film 21 is pressed against the edges of the wells 15 of the microplate, sealing and sealing the wells with the film. Further, the magnet of the magnetic unit 10b is pushed down toward the microplate membrane so that the shielding film 21 and the end of the magnet 13b contained therein are contained in the liquid 23. Thus, the microparticles 22 in the liquid 23 accumulate transversely at the end of the

Figure 30 shows an embodiment in which the magnetic units 10a and 10b do not have separate ferro-metal bushings. They are replaced by a ferro-metal plate 12 which is shaped so that the wells of the micro-plate have downwardly directed projections. The magnets 13a and *, 13b are located in the openings at the projections of the ferro-metal plate 12. In Figure 30! the magnets 13a and 13b of the magnetic units 10a and 10b are similarly in different phases. , 25 as in Figure 29.

Fig. 31 shows an embodiment in which the magnetic units 10a and 10b also have a common ferro-metal plate 12 replacing the sleeves, which in this case is a straight plate.

The magnets 13a and 13b are disposed in the openings in the ferrous metal plate 12. Also in this figure: The magnets 13a and 13b of the magnetic units 10a and 10b are in different phases. 29, unlike the solution, the protective film 21 is printed against the edges * 'of the wells of the microplate. by means of magnets 13a and 13b and not by ferro-metal bushings.

<·

The magnet 13a of the magnetic unit 10a is in the sealing position while the other 13 is in the sealing position. The magnet 13b of the magnetic unit 10b is in the collecting position of the microparticles 22.

Figure 32 shows a multi-channel transfer device 40 in which the magnetic units 10 are arranged in a circular shape. Such a device is advantageous when microparticles have to be collected. : 35 115294 for 39 large volumes. The magnetic units 10 may each have a separate protective film, but according to another embodiment, each magnetic unit 10 has one common protective film.

The above embodiments of the invention are only examples of the implementation of an idea according to the invention. It will be apparent to one skilled in the art that various embodiments of the invention may vary within the scope of the following claims.

* * ♦: · »· * · f ·» »I *» 5 · '* »» #

»I

»: T 115294 40

LIST OF REFERENCE NUMBERS

10 magnetic unit 11 rod 5 12 ferromagnetic tube or sleeve 13 magnet 14 junction 15 mouth 16 junction tube 10 17 magnetic field lines 18 magnetic field collection area 19 magnetic field 20 collection surface 21 protective film 15 22 microparticles 23 solution 24 magnetic pole 25 fluid pole 25 fluid pole 25 protective film ridge #>. 30 microparticle transfer device! 31 body tube 25 32 fitting sleeve; '33 mounting flange' · · '34 tube transfer unit 35 tube transfer pin:.' '36 elongated opening 30 37 magnet transfer plate: ·, 38 magnet transfer pin'; 39 elongated aperture 40 microparticle multichannel transfer device; * 41 magnetic unit array j 35 42 pocket 43 connecting rod 44 return spring 41 115294 45 intermediate rod 46 "triggered" 50 automatic 51 matrix 5 52 control unit 53 arrow 54 arrow 55 sample plate 56 matrix 58 (second) control unit 60 reactor unit 61 reaction vessel 62 channel 15 63 valve 64 tip section 70 cabinet 71 motor 72 actuator 20> t

Claims (12)

115294 A magnetic transfer method for sorting, collecting, transferring, or dispensing microparticles (22) or magnetic particles, either in the same liquid 5 (23) or from liquid (23a) to another (23b), by a magnetic field comprising collecting particles thereon at least one magnet (13) or the like and is dispensed by changing the magnetic field or the intensity of the magnetic field, for example by moving the magnet, characterized in that the change in magnetic field or its intensity is effected by at least one ferromagnetic body such as plate or tube one magnet (13) and / or at least one body is moved relative to one another such that when collecting the particles (22), the magnet is partially or completely outside the ferromagnetic body and when removing or dispensing particles, the magnet is partially or completely fer inside or behind a romagnetic piece. 15 Method according to Claim 1, characterized in that the strength of the magnetic field is adjusted by moving at least one magnet (13) and the ferromagnetic tube (12) relative to one another such that the magnetic field strength is reduced by moving the magnet (13) or 20 tubes (12) enters the tube, - and that the magnetic field strength is increased by moving the magnet (13), or - »·. ·. the tube (12) so that the magnet comes out of the tube. • t • · Process according to claim 1 or 2, characterized in that,. The strength of the 25 magnetic fields is reduced by moving the magnet (13) * ·; 'Inside the ferromagnetic tube (12) or by moving the ferromagnetic tube over the magnet ··'. : '4. Method according to Claim 1, 2 or 3, characterized in that, the magnetic field strength is reduced by moving the ferromagnetic tube (12) * ·. on the magnet (13) inside the hard cup shield (21) or by pushing the tube »; between the elastic protective film and the magnet. »'5. Microparticle (22) transfer device (10) for sorting, collecting, transferring or dispensing microparticles or magnetic particles: either in the same fluid (23) or from a liquid (23a) to another (23b) comprising at least one cover or protective film (21). ) internal magnet (13) or the like, characterized in that the transfer device (10) comprises at least one ferromagnetic body such as a plate or tube (12) and that at least one magnet and / or at least one ferromagnetic body such as a plate or tube (12) are movable relative to one another such that when collecting the particles, the magnet 5 is partially or wholly outside the ferromagnetic body, and when detaching or dispensing the particles, the magnet is partially or completely within or behind the ferromagnetic body. Transfer device (10) according to claim 5, characterized in that the transfer device has at least one ferromagnetic tube (12) or opening in the ferromagnetic plate and at least one permanent magnet (13) or electromagnet moving in the tube or opening, and the opening is of iron or other material whose magnetic properties prevent the magnetic flux of the magnet (13) from passing through the tube. 15 Transfer device (10) according to Claim 5 or 6, characterized in that the transfer device comprises two or more magnets (13), which are identical or different and which are attached to one another by magnetic force or by a medium or ferromagnetic or non-ferromagnetic. 20 Transfer device (10) according to Claim 5, 6 or 7, characterized in that - the magnet (13) is attached to the rod (11), by means of which the magnet can be: moved in a ferromagnetic tube (12); and that the rod (11) is ferromagnetic or non-ferromagnetic. ..,: 25 Transfer device (10) according to one of Claims 5 to 8, characterized in, '. t - that the ferromagnetic tube (12) is a circular cylinder and the magnet (13) is at least one circular rod or pin concentric with the tube, and that the magnetization axis of the magnet (13) is parallel to its longitudinal axis such that •; · 30 magnet poles are at the ends of the bar. *; Transfer device (10) according to one of Claims 5 to 8, characterized in that: - that the ferromagnetic tube (12) is a circular cylinder and the magnet (13) is at least one circular bar or pin of the same center as the tube,; 35 and that the magnet axis of the magnet (13) is transverse, i.e. perpendicular to both the ferromagnetic tube and the longitudinal axis of the rod-shaped magnet. 115294 Transfer device (10) according to one of Claims 5 to 10, characterized in that the protective film (21) is a cup-shaped material of non-stretchable material such as hard plastic or metal and that the protective film (21) forms an extension of the ferromagnetic tube (12). that the magnet (13), when pushed out of the 5 tubes, can move inside the protective film. Transfer device (10) according to one of Claims 5 to 11, characterized in that the protective film (21) is made of a stretchable and flexible material, such as an elastomeric plastic shield or a thin film, which is stretched when the magnet (13) is ejected from the ferromagnetic tube 10 (12). 115294