SE1230151A1 - Heterogeneous multi-step separation with complementary detection method - Google Patents

Abstract

The invention relates to an arrangement for separation followed by specific and ultra-sensitive detection of analytes (cells, bacteria, molecules, etc.) captured from complex environments, eg blood plasma, food extracts or seawater. The arrangement is characterized by separation in several stages: l. specifically capturing analyte on magnetic or other particles (MP) with recognition elements (eg antibodies). 2. magnetically or otherwise specifically driven transport of MP analyte complexes to detection fragments (elongated structures with specific binding sites for MP analyte complexes). The latter are bound to switch molecules (which have rapidly reversible binding to the detection fragments), for example on the wall of a narrow channel. Reversal of the bond between switch and detection conjugate followed by specifically driven transport of MP detection fragment complex within the channel to a collection site with detector. The driving force for the transport and the detection method are adapted to each other for the highest sensitivity and specificity.

Description

The detection of such a set of markers (multiplex detection) would also be desirable eg for the diagnosis of prostate cancer as isolated use of prostate-specific antigen for this purpose gives a large number of false positive results with associated discomfort for the patient and significant costs to society. Even in certain situations with infectious diseases and during inspections of food products for foodborne pathogens, it is important to detect a set of markers.

In the light of the above, it is clear that there is a great need for new technologies that are more reliable, more cost-effective and more environmentally friendly while enabling multiplex detection with high sensitivity and specificity and with the potential to be miniaturized for simplified use in portable detection systems and for analyzes even of the total protein expression of individual cells.

Biological molecular motors form the basis for the ability of living cells to move and to transport molecular and nanoscale loads between their various parts. In this context, the molecular motors interact with filamentous molecules, so-called cytoskeletal filaments (actin filaments and microtubules). Thus, a number of different variants of the molecular motor use myosin, actin filaments as interaction partners (traces that the motor travels along or fl protrudes) while the molecular motors kinesin and dynein interact with rnikrotubules. Other types of biological molecule motors are nucleic acid-based motors such as DNA and RNA polymerase that move along a nucleic acid strand. In recent years, semi-synthetic molecular motors have also developed that interact with nucleic acid strands, and there are also completely synthetic molecular motors with synthetic traces.

Characteristic of molecular motors is partly their ability to convert chemical energy (eg ATP) to mechanical energy but also their ability to alternately bind to and release from their filamentous interaction partner eg after supply of an allosteric regulator such as ATP (for myosin). The bond to the interaction partner can be switched on and off (switch molecules), for example depending on the chemical environment of the environment.

The possibilities of using molecular motors and their filamentous interaction partners (cytoskeletal filaments or nucleic acids) for applications in, for example, diagnostics have been considered for a long time (overviews in 25). Various surface chemistries that allow molecular motors to operate selectively only if they are attached along nano- or microscale grooves or channels have been studied3 '648 and enabled uniform transport along grooves / channels wß of the mentioned types of filamentary interaction partners (hereinafter referred to as filaments). Methods 2434 have also been developed for attaching recognition elements (such as biotin, antibodies, single-stranded oligonucleotides and aptamers) to said filaments or along bundles thereof (e.g. 35 "36) and then specifically binding particles, cells, molecules, etc. to the filaments via the recognition elements and transporting them on smooth or micro / nano-patterned surfaces. Prototypes for diagnostic systems också ”have also been described, in which the cytoskeletal filaments with recognition molecules first capture the analytes to be detected and then transport them to a detection area on a micro- or nano-patterned chip.

The industrial potential of using molecular motors and their cytoskeletal filaments for both separation and concentration of analytes as well as detection in practically useful equipment has become increasingly realistic in recent years in light of the advances described above and other results. The latter include the demonstration of long-term shelf life of molecular motors and cytoskeletal filaments when stored in a standard -20 ° C household freezer40 or f, .. 41 42 in other ways such as storage in the presence of a drug43 in a refrigerator or even at room temperature. The industrial applicability is also apparent from the potential improvements of diagnostic equipment that can be obtained. For example, the molecular motor-based methods enable separation without the use of pumps with associated power supplies that would otherwise be needed to drive microcirculation flows in conventional miniaturized separation equipment. The molecular motor-based systems also have advantages in being biodegradable, enabling a very high degree of miniaturization.

Finally, they can be obtained cheaply, for example from slaughterhouse waste.

Magnetic particles can be conjugated with recognition molecules (eg antibodies) that selectively bind analytes in a sample solution. Then the analytes can be separated from other molecules and particles in the solution and then detected by a variety of methods. The object of the present invention is to achieve very sensitive (low detection limit) and specific detection in a way that enables both portable equipment. detection of many analytes simultaneously (multiplex detection) and inexpensive components. We achieve this by applying the combination of switch molecules such as molecular motors, elongated detection fragments such as cytoskeletal filaments as well as nano / microparticles and magnetic, electrical or centrifugation-driven separation in a completely new way.

Solution The advances in diagnostic methods involving molecular motors and cytoskeletal filaments have to a very large extent followed a given pattern of '37 "". This has meant that the cytoskeletal filaments with recognition molecules (detection fragments) first capture the analytes to be detected and then drive of molecular motors along nanoscale or microscale channels / grooves on a surface so that the analytes are rapidly transported to a nanoscale detector on a chip.Only a few studies have concerned the capture of analyte along detection fragments in solution32 ”5355 and possible subsequent transport and separation in solution53.

What, above all, no previous study has been close to is the central point of the present invention, namely an arrangement for separation and detection with capture of an analyte from a sample solution by means of magnetic or other microparticles and accompanying step-step separation based on different specific binding processes and different selective transport steps and a suitably selected detection method where the latter is adapted to the last transport step for the highest specificity and signal / noise ratio.

Important in the first step separation is controllable and specific binding of elongated detection fragments (with a high surface-volume ratio) to a surface in one of the steps. It is further essential in the invention that the detection method and the method for the last transport step to the detector are matched to each other for the highest sensitivity and specificity.

More specifically, the present invention consists of an arrangement in which: l. An analyte in a sample solution is captured on magnetic particles (nanoparticles (<200 nm diameter), or microparticles (0.2-10 μm) or relatively large (magnetic or non-magnetic) microparticles (2-20 μm in diameter) each with one or fl your specific types of recognition molecules, 2. the magnetic particles are transported by magnetic driving forces (magnetic fields) 44 or the large microparticles without magnetic moment are transported by centrifugation or sedimentation under ordinary g- forces to many detection fragments (elongated structures with recognition molecules such as antibodies) that are reversibly and specifically bound to the surface of a channel (eg capillary) via switch molecules (which can quickly release the detection fragments at the appropriate signal), 3. magnetic particles or the the large microparticles that are not bound to the detection fragments after a certain incubation period are rinsed off56 el is removed by a suitably applied electric or magnetic field, 4. the detection fragments are then released ultra-fast (within 1-2 seconds) from the surface by a special switch signal (eg after application of a special chemical or after a light pulse or other pulse with electromagnetic radiation or sound waves), 5. complexes consisting of detection fragments with magnetic particles and analyte (s) are transported to a detector area where the amount of detection fragments and or number of magnetic particles is measured and signals the magnitude of the original analyte concentration in the sample solution. The latter transport takes place either with magnetic driving forces (when magnetic particles are used) or centrifugation-based separation or molecular motor-based transport. The transport method used here is adapted to the detection method. If molecular motor-based transport is used (requires detection fragments with actin filaments or microtubules), the presence of particles at the collection site (eg by orescence, or detection of magnetic particle 51 '57' 58) is detected for the highest specificity. In other cases, the presence of the detection fragments at the collection site is detected in order to achieve the highest specificity, for example by detecting the presence of their reporter molecules.

The invention can be adapted to multiplex detection by adequate combination of magnetic particles and detection fragments with different, suitable recognition molecules and detection fragments with different signatures / reporter molecules (for example colors).

Brief Description of the Figure Figure 1. Schematic illustration of the most important components of the invention. 1. Magnetic particles or 2. large microparticles with specific recognition elements, 3. Analyte, 4.

Detection fragments attached to 5. switch molecules on surface. 6. Scale lines corresponding to approx. 20 nm.

Figure 2. Schematic illustration exemplifying a variant of the detection arrangement where magnetic particles are used. First, samples with analyte (1) are injected into an antechamber where the analyte (2) is captured by recognition molecules attached to magnetic particles (3). The magnetic particles with bound analytes are then drawn further through a membrane (4) to a recognition chamber in the form of a channel (5) where the analyte on the magnetic particle is attracted to the walls by means of external magnets (exemplified by 6). Switch molecules (7) have been attached to the walls and, in the absence of a switch signal, they hold fixed detection fragments (8) which can now bind magnetic particles with analyte (9). When the analyte is bound, unbound magnetic particles (without analyte) are rinsed out or magnetically pulled out by the magnet (6) mot being exposed to the antechamber or in another similar way. Most often, a switch signal is given, for example, by a light pulse or that a molecule is released in the chamber or supplied from outside (10). This causes the detection fragments to detach from the switch molecules (11) and be pulled towards the other part of the chamber (12), by means of an additional external magnet (13) or by moving the magnet (6) läng along the channel towards the detection site with detector (14) . The analyte is now detected (14) by measuring how much detection fragment has been transferred to the detector, for example by registering the presence of reporter molecules on the detection fragment.

The scale bar (15) corresponds to a value between 1 μm and 5 mm depending on the exact implementation of the arrangement.

Figure 3. Schematic illustration exemplifying an arrangement with fl your parallel systems for detecting fl your analytes simultaneously (multiplex detection). To the left (1) is a cross-section illustrating that many detection chambers (2; eg capillaries) fit next to each other on a smooth surface (eg in a magnetic / conductive block). The scale bar (3) can be anywhere between 1 μm and 5 mm depending on the degree of standardization and implementation chosen for the detection arrangement. To the right is a schematic longitudinal section of a multiplex detection system (scale other than Figure 1). Each horizontal line represents a capillary extending from the atrium (4) to the detection portion (7). External (fl surface / electromagnetic) magnets are represented by (5) and (8). The middle vertical rectangle (6) represents any channel (micro-destiny) that can be used to flush out magnetic molecules that are not bound to the walls and / or to give a switch signal (eg supply certain molecules that release switch molecules from detection fragments ).

Detailed description of the invention Definitions In the context of the present application and the invention, the following definitions apply: -The term "detection fragment" denotes a polymer filament (a polymer wire) or another elongated object with a high surface / volume ratio on whose surface is fixed (covalently or otherwise) ) one or olika your different types of recognition elements that recognize one or olika your different types of analyte. The length of the detection fragment can vary between 10 nm and 1 mm and the diameter can vary between about 1 nm and 1 μm. These dimensions refer to detection fragments without recognition molecules. Examples are given below.

-The term "recognition element" or "recognition molecule" refers to anything that can be bound to the polymer filament or other elongated object and thus create a detection fragment which, with high specificity, binds to a substance (an analyte) intended to be detected in the sample. Examples are given below.

-The term "reporter molecule" is meant to mean a molecule that binds to any component in the detection kit (detection fragment, magnetic particle, recognition element) and then forms the basis for the system to detect binding of analyte to multiplex detection fragment.

The term "reporter molecule" may refer to an organic fluorophore, an enzyme or a particle such as a quantum dot or other type of nano- or microparticle. Further examples are given below.

The term "switch molecule" refers to a molecule that binds a detection fragment specifically and reversibly and with very fast kinetics. By fast kinetics is meant here that the switch molecule, after a "switch signal" in the form of a light pulse or supply of a certain molecule (a certain ion, adenosine triphosphate (ATP) etc.) releases the detection fragment very quickly (in less than 1- 2 s). By specific binding between detection fragment and switch molecule is meant that the switch molecule only binds strongly to the given detection fragment and not to other components in the detection arrangement such as analyte, microparticle, etc. Examples are given below.

-The term "switch signal" refers to a signal (light pulse, supply of a certain nucleus, etc.) that releases the detection fragment from the switch molecules ultra-fast, within <2 s. Examples are given below.

The term "analyte" is intended to mean a substance or component that a person, knowledgeable in the field, would wish to detect. Examples are given below.

-The term "condition to be detected" is meant to mean a condition (for example, a particular disease state or the presence of a particular food pathogen in a sample) that a person skilled in the art would wish to detect. Examples are given below.

Schematic illustration of recognition molecule, detection fragment and switch molecules is given in Figure 1.

The invention aims at detecting one or more analytes in a sample by means of specific recognition and a completely new combination of detection method (s) and olika different separation steps in series. As a number of concepts appear frequently in the description below, the presentation begins by giving examples of these. They have otherwise been more precisely and generally defined above.

Samples or sample solutions of interest in which different analytes are to be fed are selected from groups consisting of water (seawater, lake water, drinking water or other), soil, food components, air and biological samples such as blood, blood plasma, blood serum and other body fluids (such as tears, saliva , secretions, cerebrospinal fluid, urine) or extracts thereof, cell or tissue extracts, extracts of feces from birds and other species, etc.

The analytes to be detected may belong to the group of biomarkers, eg certain proteins, oligonucleotides or DNA or RNA fragments. They may also belong to groups such as bioterrorism threats, environmental risks and / or various pathogens (viruses, bacteria, parasites, etc.), for example of relevance to human health, animal health, plant health or food safety.

"Conditions to be detected" include the presence of harmful substances / organisms in samples or sample solutions of interest, the presence of specific biomarkers for various diseases or sub-groups of diseases (eg various cancers such as breast cancer, prostate cancer, leukemia; various degenerative nerve diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis; various autoimmune diseases such as rheumatoid arthritis, multiple sclerosis; diseases such as stroke and myocardial infarction) in samples or test solutions and the presence of markers of inflammation such as C-reactive protein in samples or sample solutions.

Examples of recognition elements are monoclonal antibodies, polyclonal antibodies, Fab fragments, chimeric proteins, lectins, receptor proteins, nucleic acids, oligonucleotides, aptamers, peptides, chemical substances or fragments thereof.

These recognition elements are attached both to magnetic particles and to what becomes a detection fragment in such a way that the magnetic particles and the detection fragments recognize different parts of an analyte, whether it is a cell, particle, molecule or other.

The detection fragment consists of unmodified, or genetically modified or otherwise engineered protein filaments (amyloid fibrils, actin filaments, microtubules, including their prokaryotic analogs and fragments and chemically modified versions of them, etc.) or synthetic or serni-synthetic polymer filaments, nanowires or nanotubes or nanotires loaded with recognition elements (see above) against an analyte or set of relevant analytes whose concentration (s) are expected to change to a certain level (either all increased or all decreased or certain increasing and other decreasing) under a condition intended to be detected, e.g. a particular disease or a sub-type of a disease. The detection fragment has also bound reporter molecules and can be released from a surface of switch molecules. For dimensions of detection fragments, see above.

Examples of reporter molecules are colors; fl uorophores; enzymes (eg horseradish peroxidase, HRP; alkaline phosphatase); quantum dots; nanoparticles of gold, silver or other metal or material; nucleic acids; oligonucleotides; mm.

As mentioned above, the switch molecules reversibly bind to the detection fragments.

The switch molecules must therefore be adapted to the later fragments for specific and reversible binding controlled by a switch signal. Molecular motors are switching molecules for binding cytoskeletal filaments or DNA strands. Properly functionalized G proteins and molecular motors can also function as switch molecules if they are provided with a structural moiety that specifically binds to the detection fragments under certain conditions but where this binding can be reversed by a switch signal. Other molecules that change their structure during illumination or when a certain chemical is added can also function as switch molecules in combination with suitable detection fragments.

A switch signal can be a pulse of electromagnetic or other radiation (eg light or ultrasound), the supply of a chemical (eg adenosine triphosphate; ATP) or a combination of your chemicals (eg ATP and a high (or low) concentration of salt ions such as sodium, potassium, chloride, etc.) or altered pH or temperature. These changes can be induced in different ways. For example, ATP can be generated within the channel with detection fragments through activated enzymatic activity, or released from so-called caged-ATP59 or from other complexes. Chemicals that act as switch signals can also be released from containers (liposomes, vesicles, polymer containers or other) 60 inside the channel.

The invention in its entirety: The present invention consists of an arrangement where: 1. A sample solution is supplied to an antechamber (Figure 2). There, analyte or analytes from the sample solution are captured on magnetic particles (nano (<200 nm diameter), or microparticles (0.2-10 μm) or relatively large (magnetic or non-magnetic) microparticles (2-20 μm in diameter) each with one or fl your specific types of recognition molecules.

The recognition molecules have been adsorbed to said particles or covalently bound to them or via biotin-streptavidin or other linkers. 2. the magnetic particles are transported directed by magnetic driving forces (magnetic fields) 44 or the large microparticles without magnetic moment are transported by means of electric driving forces, centrifugation or sedimentation under ordinary g-forces to many detection fragments that are reversibly and specifically bound to the surface / wall of a recognition chamber, eg a channel (eg capillary; of glass, plastic, metal or other) via switch molecules. The magnetic transport is driven by either surface permanent magnets or stationary or surface electromagnets. The channel can be made of glass, plastic, metal or other material. 3. magnetic particles or the large microparticles which are not bound to the detection fragments after a certain incubation period are washed away56 or transported away by magnetic driving forces eg by turning an electromagnet on or off, moving one or fl your permanent magnets or by electric driving forces (eg electrophoresis due to electrostatic forces or dielectrophoresis due to oscillating electric fields) or capillary force. 4. the detection fragments are then released ultra-fast (within 1-2 seconds) from the surface by a special switch signal (eg after application of a special chemical or after a light pulse or other pulse with electromagnetic radiation or sound waves; see above). A new chemical can be added, for example, by electroosmotic fate or by diffusion from a container. 5. complexes consisting of, among other things, detection fragments with magnetic particles and analyte (s) are transported to a detector area / collection point where the amount of detection fragments and or number of magnetic particles is measured and signals the magnitude of the original analyte concentration in the sample solution. The latter transport takes place either with magnetic driving forces (when magnetic particles are used) or centrifugation-based separation or molecular motor-based transport. The transport method used here is adapted to the detection method. If molecular motor-based transport is used (requires detection fragments with actin filaments or microtubules), the presence of particles at the collection point (eg by orescence, or detection of magnetic particles "57" 58) is detected for the highest specificity. In other cases, the detection fragments are detected, for example by detecting the presence of their reporter molecules, for example by orescence, formation of colored substance or free electrons by enzymatic reaction catalyzed by enzyme such as horseradish peroxidase (HRP) or alkaline phospholmate as alkaline phosphorate to the detection fragment.

The variant of the invention which utilizes magnetic particles and magnetic fields as driving forces as well as detection of the detection fragment is schematically illustrated in Figure 2.

Further development for multiplex detection: Multiplex detection can be made possible in a number of different ways: l. By using fl your parallel channels (Figure 3) with molecular motors and detection fragments as above. In each of these, detection fragments have then been bound (to switch molecules) which recognize a certain analyte and which are labeled with a certain reporter molecule. Alternatively or simultaneously, each channel may have olika your different types of detection fragments with recognition elements for analy your analytes and different reporter molecules.

Each parallel channel is connected to a container with magnetic particles with recognition elements against another part / other parts of the same analyte (s) that detection fragments in the channel recognize. Further multiplexing can be achieved using a) multiplexing detection fragments where each detection fragment has a set of different recognition elements bound to the fragment and b) a mixture of magnetic particles with recognition elements bound to them, recognizing the same molecules as the recognition elements on the detection fragments.

Nvhet and inventive step The present invention has not been previously described. Magnetic separation has been described (see above) but not combined with separation based on detection fragments or the equivalent reversibly and specifically bound to switch molecules on a surface. If detection fragments have been used to capture analytes on a surface (as for the present invention), they have not done so in a channel (such as a cannula or a capillary) but on a nano- or micro-patterned surface where molecular motors then transported detection fragments to a nanoscale detector surface37. Furthermore, the release of complexes consisting of, for example, detection fragments with cross-linked magnetic particles (which occurs here when switch molecules are affected by sWitch signal) has not been accompanied by magnetic or other non-molecular motor-driven transport of this entire complex to a detector. Given the rest of the research and technology field's current focus on molecular motor-based transport on a chip after binding of analyte to the detection fragment, it is unlikely that a group of people with adequate technical competence would come up with the idea described here. The first research group that described molecular motor-based transport of model analytes to a detector area on a chip37 has also described electrophoresis-based transport of cytoskeletal filaments to a detector area in a nanocanal53. However, the electrophoretic transport in that study lacked the specificity that is the great strength of the arrangement described here. This is because the electrophoretic transport53 of cytoskeletal filaments transported all filaments to the detector whether or not they bound analyte. The high specificity of the current method is due to the fact that only the detection fragments that bound magnetic particle or large microparticle via analyte are transported by magnetic forces or centrifugal forces / gravitational forces to the collection area / detector area. Furthermore, in the present arrangement, a detector is selected which detects the reporter molecules on the detection fragments which are only transported to the detector if they are bound to magnetic particle or large microparticle because the driving force for the transport acts on them. Finally, electrophoretic transport has not previously been combined with specific and reversible binding of molecular motors to the cytoskeletal filament. The fact that world-leading innovative researchers in the field (see above), who have studied both molecular motor-based transport on a chip (to a detector surface; an example of a specific recognition between detection fragment and switch molecule) and transport of detection fragments within a channel (said electrophoresis ), without combining the two systems into a much more sensitive and specific one as described here, is very strong support for the height of the invention.

The above reasoning for the combination of magnetic particle and detection fragment applies analogously to the combination of detection fragment and large microparticles with recognition elements if the magnetic driving force is replaced by centrifugation or gravitational forces or electrostatic fields and the particles are positively charged, for example.

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Claims (20)

REQUIREMENTS Heterogeneous step separation with complementary detection method 1. Arrangements for separation and detection with capture on particles of an analyte (eg molecule, microorganism / pathogen) from a sample solution and accompanying fl step separation based on different types of specific binding processes and different selective transport steps and a suitably selected detection method where the latter adapted to the last transport step for maximum specificity and signal / noise ratio. Arrangement according to claim 1, wherein the analyte is captured on micro- or nanoparticles with suitable recognition elements. Arrangement according to claims 1 and 2, wherein the analytes after being captured by the recognition elements on the particles according to claim 2 are transported in a directed manner (eg with magnetic driving forces, electric driving forces, capillary force, centrifugation or gravitational forces) to elongated detection fragments reversibly bound to the switch molecules. of a channel of glass, plastic, metal or other material. Arrangement according to claims 1 to 3, wherein the recognition elements are taken from one or more of the groups: monoclonal antibodies, polyclonal antibodies, Fab fragments, chimeric proteins, lectins, receptor proteins, nucleic acids, oligonucleotides, aptamers, peptides, chemical substances or fragments of these. Arrangement according to claims 1 to 4, wherein the detection fragments consist of unmodified, or genetically modified or otherwise manipulated protein filaments (amyloid fibrils, actin filaments, microtubules), including their prokaryotic analogs and fragments and chemically modified versions of them, etc.) or synthetic or semi synthetic polymer filaments or nanowires, each loaded with recognition elements (see above) against an analyte or set of relevant analytes (enabling multiplex detection) whose concentration (s) are expected to change to a certain level (either all increased or all decreased or some increasing and other decreasing) during a condition that is intended to be detected, eg a certain disease or a sub-type of a disease. Arrangement according to claims 1, 3 and 5, wherein reporter molecules are attached to the detection fragments. Arrangements according to claims 1,3, 5 and 6, wherein reporter molecules are colors; fl uorophores; enzymes (eg horseradish peroxidase, HRP; alkaline phosphatase); quantum dots; nanoparticles of gold, silver or other metal or material; nucleic acids; oligonucleotides; mm. Arrangements according to claims 1, 3, and 5, wherein switch molecules are constituted by molecular motors such as myosin, kinesin, G-proteins or other molecules or nanostructures which change their structure in response to changing the environmental conditions in such a way that they break their specific binding to the detection fragments. Arrangement according to claims 1-5, wherein the analytes captured on particles, after transport, are allowed to bind to detection fragments during a certain incubation period, after which the particles not bound to detection fragments are washed away or transported away (eg with magnetic driving forces, electric driving forces , capillary force, centrifugation or gravitational forces). Arrangement according to claims 1-6 and 9, wherein the detection fragments are released from switch molecules by a special switch signal. Arrangement according to claims 1-6 and 9-10, wherein the switch signal relates to the supply of special chemical (s) / molecule (s) or a combination of special chemicals / molecules and / or a light pulse or other pulse with electromagnetic radiation or sound waves or changed temperature, pH or other. Arrangement according to claims 1-11, where complexes consisting of detection fragments with bound analytes and particles, in the last transport step before detection, are transported in a directed and specific manner (eg by means of molecular motors or magnetic driving forces, electric driving forces, capillary force, centrifugation or gravitational forces). to a detector area and where the transport method is adapted to the detection method so that what is detected is not essential for the last transport step. Arrangement according to claims 1-12, wherein the presence of the detector fragment reporter molecules or other specific properties of the detection fragment is detected by the detector, for example by fl uorescence or otherwise, in cases where the last transport step according to claim 12 is caused by magnetic driving forces, electric driving forces. capillary force, centrifugation or gravitational forces. Arrangement according to claims 1-12, wherein the presence of particles as magnetic particles or large microparticles bound to the detection fragment (rather than the detection fragment itself) is sensed by, for example, fl uorescence or otherwise in the case the last transport step according to claim 12 is caused by molecular motors driven detection fragment with particles to the detector site. A "kit" consisting of particles with recognition elements, detection fragments and an apparatus with a channel, magnets, and all the equipment needed for detection etc. and a manual for the use of said particles, detection fragments and apparatus. A "kit" according to claim 15, wherein multiplex detection (detection of several analytes simultaneously) is made possible by the apparatus having fl your channels each with detection fragments with recognition elements against one or fl your analytes and where these are associated with suitable sets of magnetic or other particles. with different recognition elements. A method of detecting analyte in a sample comprising: a. Providing a sample b. Supplying a sample to a detection fragment according to any one of claims and c. Detecting said analyte A method according to claim 1-17, wherein analyte is detected by detecting magnetism and / or ores uorescence from particles, absorbance of light, light scattering or amperometric recording and / or detecting the presence of reporter molecules in detection fragments by fl uorescence, absorbance of light, light scattering, magnetism , amperometric or other registration. 19. l9. Method according to claims 1-18 where the sample provided can be water (sea water, lake water, drinking water or other), soil, food components, air and biological samples such as blood, other body fluids (such as tears, saliva, secretions, cerebrospinal fluid, urine) or extracts thereof, cell or tissue extracts, extracts of feces from birds and other species, etc. Use of particles with recognition elements, detection fragments, "kit" or the detection method for the detection of one or more analytes.