WO1997042503A1 - Procede et installations de separation de particules magnetiques dans un fluide pour l'analyse biologique, et application dudit procede - Google Patents
Procede et installations de separation de particules magnetiques dans un fluide pour l'analyse biologique, et application dudit procede Download PDFInfo
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- WO1997042503A1 WO1997042503A1 PCT/FR1997/000794 FR9700794W WO9742503A1 WO 1997042503 A1 WO1997042503 A1 WO 1997042503A1 FR 9700794 W FR9700794 W FR 9700794W WO 9742503 A1 WO9742503 A1 WO 9742503A1
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- cells
- tube
- magnetic
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
- G01N1/405—Concentrating samples by adsorption or absorption
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
- B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
- B03C1/00—Magnetic separation
- B03C1/02—Magnetic separation acting directly on the substance being separated
- B03C1/035—Open gradient magnetic separators, i.e. separators in which the gap is unobstructed, characterised by the configuration of the gap
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54326—Magnetic particles
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N35/0098—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S435/00—Chemistry: molecular biology and microbiology
- Y10S435/971—Capture of complex after antigen-antibody reaction
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S436/00—Chemistry: analytical and immunological testing
- Y10S436/806—Electrical property or magnetic property
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S436/00—Chemistry: analytical and immunological testing
- Y10S436/807—Apparatus included in process claim, e.g. physical support structures
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S436/00—Chemistry: analytical and immunological testing
- Y10S436/807—Apparatus included in process claim, e.g. physical support structures
- Y10S436/81—Tube, bottle, or dipstick
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S436/00—Chemistry: analytical and immunological testing
- Y10S436/824—Immunological separation techniques
Definitions
- the present invention relates to a method and an installation for separating magnetic particles in a fluid for the biological analysis of rare events.
- the invention more particularly finds its application in the fields of medical diagnosis and quality control, in particular in the food industry, but also in the therapeutic field for detecting and taking from samples a cell type with a low occurrence.
- medical applications of the invention there may be mentioned:
- Separation of fetal cells present in maternal blood involves selectively collecting cells present at the rate of approximately 1 fetal cell per 10 million non-fetal cells.
- the implementation of the present invention should make it possible to replace the risk samples of amniotic cells.
- immuno-affinity methods consisting in fixing an antibody on a support, which antibody reacts vis-à-vis an antigenic motif present on the surface of the cells sought (Forsgren and Sjoquist, 1986; Langone, 1982).
- FACS Fluorescence Activated Cell Sorting
- MCS Magnetic Affinity Cell Sorting
- FACS and MACS methods are well known today and have already given rise to industrial implementation. In general, the results obtained with each of these methods differ little in terms of effectiveness and allow between 70 and 100% recovery. However, they are more or less suitable for the various applications envisaged.
- the present invention is also based on the use of magnetic particles on the surface of which is immobilized a substance capable of specifically binding to a complementary substance which is free or present on the surface of cells.
- the analysis is carried out in a liquid medium by subjecting the biological medium into which said magnetic particles have been introduced to a magnetic field.
- the magnetic particles more particularly envisaged within the framework of the invention are microbeads of the type of those sold by the companies Dynal, Rhône Poulenc or Sigma.
- paramagnetic microbeads of micron size can have on their surface different ligands depending on the application which is envisaged. These ligands are fixed to the surface of the microbeads by the techniques described in the prior art, these ligands can be:
- Oligonuleotides for example oligo (dT) or oligo (dA) of variable sizes, biotinylated and therefore fixed to the surface of the microbeads by means of streptavidin.
- oligonucleotides make it possible, by hybridization, to purify or extract RNA or DNA from samples previously treated so as to render the nucleic acids which it contains capable of hybridizing.
- the use of these oligonucleotides attached to the surface of the paramagnetic beads can also constitute a preparatory phase for an amplification protocol by PCR or a cloning after PCR, or even be used for sequencing in solid phase.
- the beads thus loaded with a ligand are brought into contact with the sample which may have undergone a preliminary treatment, in order to achieve the segregation of the event sought by the action of a magnetic field.
- the use of this technique has the drawback of requiring multiple manipulations and leads to a set of beads more or less aggregated by the action of the magnet, among which are some rare cells.
- the disproportion of the number of beads compared to the number of cells, of the order of 1 to 10 then makes the identification and collection of these cells impossible.
- the object of the present invention is therefore to offer a method eliminating the steps of rinsing and elimination of cells and liquids not sought and jointly the automatic selection of the sought elements and the segregation with free paramagnetic beads.
- Another object of the present invention is to improve the efficiency of the magnetic methods by avoiding the artefacts noted in the methods of the prior art and by simplifying the implementation protocol.
- the sensitivities sought in the abovementioned fields can be of the order of 1 cell for 1 or even 10 grams of sample.
- the European patent EP206077 is also known in the state of the art.
- This patent discloses the general principle of magnetic immunoseparation by circulation in a tubular segment placed in a uniform magnetic field. It has appeared in certain use cases that the excess particles could be attracted too quickly at the start of the tube, and that agglomerates were created blocking the circulation of the labeled or unmarked cells. Given the scarcity of target cells, this situation is very troublesome.
- the invention aims to remedy this drawback by proposing to create an inhomogeneous magnetic field. This characteristic prevents the formation of agglomerates of unbound particles in the upstream part of the tubular segment.
- the invention firstly relates to a method of magnetic separation of cells, in particular bacteria, fetal cells, stem cells of the bone marrow and of circulating cancerous cells, of the type consisting in fixing the target cells. on paramagnetic beads and causing a magnetic field to act on a sample containing the fixed cells, the free cells and the excess paramagnetic beads, to isolate the paramagnetic beads, characterized in that the sample is circulated in a tube whose the section is much less than the length over which the magnetic field is applied.
- the sample preferably flows at a speed of a few centimeters per second.
- the excess beads due to their lower density than that of marked or unlabeled cells, or the clusters of paramagnetic beads, due to their high sensitivity to the magnetic field, are quickly attracted against the wall of the tube and are immobilized in the part proximal of the tube.
- Unfixed cells are insensitive to the magnetic field and pass through the tube.
- the cells fixed on paramagnetic beads are entrained by the flow inside the tube, before being immobilized against the wall of the tube, in the distal part of the tube.
- the sample is made to circulate in a spirally wound tube placed in a magnetic field whose lines of flows are substantially perpendicular to the plane of the spiral.
- the section of the tube is between 0.5 and 3 millimeters and the length of the tube is greater than 10 centimeters.
- the tube is placed in a non-homogeneous magnetic field.
- This field can be produced by a permanent magnet or by an electromagnet.
- the preparation is introduced into a reservoir connected to the separation tube, said reservoir being raised relative to the tube to ensure circulation of the sample by gravity.
- the invention also relates to an installation for the magnetic immuno-separation of cells, in particular bacteria, fetal cells, stem cells of the bone marrow and of circulating cancerous cells, characterized in that it consists of a tube whose cross section is much less than the length, said tube being wound to form a spiral, and by at least one magnet arranged to create a magnetic field substantially perpendicular to the plane of the spiral formed by said tube.
- it comprises a tube having two sections of increasing section, the second section being at least 10 times greater than the first section.
- the invention also relates to applications of this method for:
- FIG. 1 shows a front view of a device according to the invention
- Figure 2 shows a sectional view of the tubing;
- Figure 3 shows a view of an alternative embodiment.
- FIGS. 6 shows a cross-sectional view of an alternative embodiment
- Figure 5 shows a view of a second alternative embodiment using a peristatic pump
- Figures 6 shows a view of another alternative embodiment in operates a peristatic pump
- FIGS. 6 ' represents a view of another alternative embodiment using a peristatic pump
- FIG. 1 represents a front view of an exemplary embodiment of a device according to the invention. It consists of a receptacle (1) which can contain approximately 10 to 50 milliliters, connected to a tube (2) wound in a spiral and ending in an outlet tube (3) provided with a valve (4).
- a set of permanent magnets (5) is arranged radially to create a non-homogeneous magnetic field. The arrangement of the magnets is indicated by way of example, a different arrangement, for example in the form of parallel magnets, also being applicable.
- the magnets (5) consist of bars made of samarium-cobalt. These magnetic bars can optionally be mounted on a rotating support disc.
- This variant makes it possible to drive the paramagnetic balls fixed towards the outlet of the tubing at one stage of the process. For example, they rotate alternately ⁇ 20 °.
- the maximum drive speed is a few millimeters per second, so as to optimize the efficiency on paramagnetic particles.
- the particles will be mixed to facilitate rinsing and the elimination of cells and particles.
- the cross-section of the tubing (2) is approximately 0.5 millimeters. It is made of a material such as TEFLON (registered trademark). The length is about 50 centimeters.
- the use of the device is as follows: a) first of all a PBS (commercial name) type rinsing liquid is introduced into a 20 milliliter pipette; b) the blood sample (7) which has been previously incubated with paramagnetic particles is aspirated. 9 milliliters of physiological saline are added (8). c) the receptacle (1) is connected to the spiral tube (2), scrupulously avoiding the formation of bubbles; d) the receptacle (1) is raised by about 30 centimeters relative to the spiral e) the valve (6) provided between the receptacle (1) and the tube (2) is opened.
- a PBS commercial name
- the next step consists in recovering the rosettes or nuclei from the cells sought.
- the spiral (2) is moved away from the influence of the magnetic bars (5), and the flow of the rest of the rinsing liquid is resumed. This one will entrain all the paramagnetic particles and the cells carrying paramagnetic particles.
- the desired particles can thus be recovered by filtering, using a filter allowing the paramagnetic particles to pass, but not the cells.
- a new reagent will be activated which will have the property of lysing the fixed cells by releasing the hardened and colored nuclei.
- a reagent consists of the following mixture:
- CETRIMIDE Trademark
- This mixture is introduced into the receptacle (1) so that the volume does not exceed that of the spiral (2).
- the coloring makes it easier to visualize the progression of the liquid.
- This mixture is left in contact for 10 minutes with the spiral.
- the magnets are actuated in alternating movements to facilitate the release of the nuclei. We can then rinse to recover the pure nuclei.
- the magnets (5) are always present, no magnetic particles will contaminate the preparation.
- the identification of the nuclei implies the use of molecular hybridization reagents. These make it possible to detect genetic anomalies such as the presence of a supernumerary centromere (trisomy 21) or amplified expressions of oncogenes, or even expressions of particular genes type P53.
- the stoichiometrically colored pure nuclei make it possible to define a level of ploid which characterizes the stages of tumor cells.
- Figure 1 shows an alternative embodiment of a device according to the invention.
- the device comprises three containers (31 to 33) of reagents and samples connected to a separator pipe (35) spirally wound. Automatic taps or valves open or close the flow of each of the containers (31 to 33) to the separator pipe (35).
- This separator pipe is placed in a magnetic field gradient generated by permanent magnets (36) arranged in a star, in alternating radial direction.
- the downstream end of the separator pipe (35) opens on the one hand into a collection pipe (37) and on the other hand into a discharge pipe (38).
- Valves or taps are provided on each of these pipes to control the flow from under to a rejection container (39), under to analysis means formed by a collection system on filter (40), by a collection container (41) or by a particle flow detector (42).
- Figure 2 shows a sectional view of a segment of tubing. Under the effect of Bernoulli's forces, the particles tend to concentrate in the center of the flux, where speed is most important. Under the effect of magnetic fluxes, the paramagnetic beads (10) lighter and less bulky than the unmarked particles (11) than the marked cells (12) are very quickly attracted to the wall (13) of the tubing.
- the trajectory of the unmarked cells (11) is not disturbed by the magnetic fluxes and these therefore remain in the center of the flux where they are quickly drawn towards the exit.
- the marked cells (12) are found in the last part of the tubing (2).
- the tubing can be made by two segments of increasing section, the first serving to retain the excess beads, and the second to retain the marked cells.
- Figures 3 and 4 show views respectively in median section and in cross section of an alternative embodiment.
- the tubing (2) is formed by a groove (14) made in a plastic plate (15). It opens on the one hand on one of the side edges to allow connection to a connecting pipe with the receptacle (1) supply, and on the other hand with a transverse orifice (16) passing through the plate (15) allowing the evacuation of fluids.
- the device is formed of two symmetrical plates (15, 15 ') and are connected so as to define between them the liquid circulation pipe. They have housings for the magnets (17) on the exterior surfaces.
- FIG. 5 represents a view of a second alternative embodiment using a peristatic pump.
- the progression of the liquids is not conditioned by gravity, but by a peristatic pump.
- the device consists of a tubular element (22) forming a separating loop. One end sucks the treated sample as in the previous examples.
- the tubular element (22) is arranged in the field of permanent magnets (24 to 28) oriented perpendicular to the direction of movement of the liquid, in alternating directions.
- the tubular element (22) is disposable, which avoids any contamination of its interior walls.
- This tubular element (22) is connected to the suction tube of a peristaltic pump (29) of known type.
- the waste is collected in a test tube (30) at the outlet of the pump (29).
- the circulation of the fluid inside the tube is preferably continuous, with a speed of one order of 1 cm per second. The continuous circulation prevents the formation of cellular adhesions on the wall of the separation tube.
- FIGS. 6, 6 and 6 show a partial view of an alternative embodiment.
- the magnets form a squirrel cage (32) formed by a plurality of magnetic bars (33 to 40) magnetized radially in alternating directions.
- This structure allows one or more tubular elements (22) to be positioned and several samples to be processed in parallel.
- the use of such a device is as follows:
- the free end (23) of the separator pipe (22) is immersed in the sample which has previously received the paramagnetic particles.
- It activates the pump (29) which will pass the magnetic particles and cells in the separator loop arranged in the magnetic field.
- the magnetic particles and the rosettes formed by the marked cells will stop in this loop while the other cells will evacuate.
- it is preferable to replace the peristaltic pump by another means of aspiration, for example a syringe suction or a source of depression, to avoid crushing of unmarked cells.
- Image analysis is able to resolve certain discriminations.
- the total number of artifacts must remain below these values, which requires high performance of the chain of separation of marked cells and unlabeled cells, and high quality of cell recovery.
- the usual slide collection and staining procedures generate "noise" which is not compatible with such requirements.
- the filtration device according to the invention makes it possible to meet these requirements. It consists of a piece (41) of molded silicone, for single use. This part (41) has a thickness of the order of 2 millimeters and has twelve protrusions (51) with a diameter of 6 millimeters and a height of 10 millimeters. These protrusions (51), normally closed, can be perforated by a hollow needle (48) provided at the end of the separator tube (42).
- the silicone part (41) is positioned in a rigid perforated plate (43).
- the perforations are arranged in a manner complementary to the protuberances (51) of the part (41).
- This plate (43) can be tightly connected to a support (45) and held in position by claws (44).
- An O-ring completes the sealing of this assembly.
- the support (45) also has perforations arranged opposite the perforations of the plate (43).
- the support is connected to a vacuum source.
- a microporous filtration membrane (47) is disposed between the plate (43) and the support (45). It is advantageously a microporous polycarbonate membrane whose cells have a section of 2 microns.
- the part (41) is delivered with a protective film placed under the underside. One or more protrusions can be opened.
- the plate (41) is placed on the support (43) after positioning the membrane microporous, and the assembly is locked using the claws (44).
- the separation tube (2) is delivered with a protective cap which prevents any contamination. This cap is removed and one of the protuberances of the part (41) is perforated.
- the isolated paramagnetic particles pass through the membrane while the rosettes remain on the surface.
- the porosity of the microporous membrane allows the passage of paramagnetic particles whose cross-section is of the order of a micron, but not the rosettes, whose cross-section is of the order of 5 microns.
- the porosity can be 1 to 2 microns.
- the coloring can be done before filtration by fluorescent dyes of the IP or BET type (trade names) or even by coloring on the filter after the latter has been extracted from the containment system.
- the combination of a separator device formed by a tube of section much less than the length placed in a magnetic field gradient, and a filtration system as described above, has many advantages. Such an association first of all facilitates the automation of biological analysis. It also makes it possible to carry out analyzes and uses for therapeutic purposes requiring a high degree of sterility.
- the paramagnetic beads allow sorting of the cells, and in particular a separation of the sought cells and the other cells.
- the filter removes certain excess cells, such as platelets, as well as excess paramagnetic beads.
- the filter also has the function of cell concentration rare sought on a reduced surface facilitating the analysis by imagery. This filter also makes it possible, for therapeutic applications in particular, to carry out a culture of the cells sought.
- the filter serves as a cell support and can be supplied with a nutritive liquid capable of promoting cell multiplication.
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- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Hematology (AREA)
- General Physics & Mathematics (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- Analytical Chemistry (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
Claims
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP9539592A JP2000500871A (ja) | 1996-05-07 | 1997-05-05 | 生物学的分析用流体内の磁気粒子の分離方法及び設備ならびにその方法の応用 |
EP97923133A EP0855030A1 (fr) | 1996-05-07 | 1997-05-05 | Procede et installations de separation de particules magnetiques dans un fluide pour l'analyse biologique, et application dudit procede |
US08/952,493 US6143577A (en) | 1996-05-07 | 1997-05-05 | Process and installations for separation of magnetic particles in a fluid for biological analysis, and application of said process |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
FR9605727A FR2748569B1 (fr) | 1996-05-07 | 1996-05-07 | Procede et installation de separation de particules magnetiques dans un fluide pour l'analyse biologique, et application dudit procede |
FR96/05727 | 1996-05-07 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1997042503A1 true WO1997042503A1 (fr) | 1997-11-13 |
Family
ID=9491932
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/FR1997/000794 WO1997042503A1 (fr) | 1996-05-07 | 1997-05-05 | Procede et installations de separation de particules magnetiques dans un fluide pour l'analyse biologique, et application dudit procede |
Country Status (5)
Country | Link |
---|---|
US (1) | US6143577A (fr) |
EP (1) | EP0855030A1 (fr) |
JP (1) | JP2000500871A (fr) |
FR (1) | FR2748569B1 (fr) |
WO (1) | WO1997042503A1 (fr) |
Cited By (1)
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US7651838B2 (en) | 2001-04-30 | 2010-01-26 | Institut National De La Sante Et De La Recherche Medicale | Prenatal diagnosis method on isolated foetal cell of maternal blood |
Families Citing this family (23)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP3551293B2 (ja) * | 1998-02-02 | 2004-08-04 | 東洋紡績株式会社 | 核酸抽出装置 |
CA2370215C (fr) * | 1999-05-04 | 2006-01-31 | Dan A. Pankowsky | Produits et methodes utilisant un parametre unique ou plusieurs parametres pour determiner des phenotypes de cellules |
US20030095897A1 (en) * | 2001-08-31 | 2003-05-22 | Grate Jay W. | Flow-controlled magnetic particle manipulation |
EP1687323A4 (fr) | 2003-08-08 | 2009-08-12 | Life Technologies Corp | Methodes et compositions pour clonage direct de molecules d'acides nucleiques |
FI20040159A0 (fi) | 2003-10-20 | 2004-02-02 | Bio Mobile Oy | Magneettinen siirtomenetelmä, mikropartikkelien siirtolaite, ja reaktioyksikkö |
US7042322B2 (en) * | 2004-04-02 | 2006-05-09 | Chien-Hsing Li | High-performance liquid magnetizer |
US7781228B2 (en) * | 2005-04-07 | 2010-08-24 | Menon & Associates, Inc. | Magnetic resonance system and method to detect and confirm analytes |
US7985340B2 (en) * | 2005-12-02 | 2011-07-26 | Invitrogen Dynal As | Magnetic separator |
WO2007063529A2 (fr) | 2005-12-02 | 2007-06-07 | Invitrogen Dynal As | Separateur magnetique |
FI20051248L (fi) | 2005-12-02 | 2007-06-03 | Bio Nobile Oy | Biologisten komponenttien rikastusyksikkö ja rikastusmenetelmä |
US20070166730A1 (en) * | 2006-01-19 | 2007-07-19 | Menon & Associates, Inc. | Magnetic resonance system and method to detect and confirm analytes |
US8304203B2 (en) * | 2007-12-05 | 2012-11-06 | Zyomyx, Inc. | Cell assay kit and method |
US20090321337A1 (en) * | 2008-06-25 | 2009-12-31 | Robert Kersten | Water vitalizing system, apparatus, and method therefor |
EP2421654A1 (fr) * | 2009-04-22 | 2012-02-29 | Clinical Genomics Pty Ltd | Procédé et appareil pour isoler une entité biologique cible à partir d'un échantillon biologique |
JP5846609B2 (ja) | 2010-01-21 | 2016-01-20 | バイオセップ リミテッド | 希少細胞の磁気分離 |
DE102011088741B4 (de) | 2011-12-15 | 2013-07-25 | Institut für Bioprozess- und Analysenmesstechnik e.V. | Verfahren und Anordnung zum Markieren und Separieren von Zellen aus einer Zellsuspension |
EP2955521A1 (fr) * | 2014-06-11 | 2015-12-16 | Centre Hospitalier Universitaire Vaudois (CHUV) | Procédés pour séparer des cellules |
CN105624035A (zh) * | 2016-02-04 | 2016-06-01 | 关节动力安达(天津)生物科技有限公司 | 基于螺旋状毛细管的细胞磁分选装置及细胞磁分选装置方法 |
EP3238759B1 (fr) | 2016-04-29 | 2019-07-17 | Fenwal, Inc. | Système et procédé de traitement, d'incubation et/ou de sélection de cellules biologiques |
US10449283B2 (en) | 2016-04-29 | 2019-10-22 | Fenwal, Inc. | System and method for selecting and culturing cells |
US10274495B2 (en) | 2016-12-21 | 2019-04-30 | Fenwal, Inc. | System and method for separating cells incorporating magnetic separation |
JP2019049455A (ja) * | 2017-09-08 | 2019-03-28 | 東芝テック株式会社 | 試料調製装置及び試料調製方法 |
JP2019158766A (ja) * | 2018-03-15 | 2019-09-19 | 東芝テック株式会社 | 濾材及び試料調製装置 |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0206077A2 (fr) * | 1985-06-22 | 1986-12-30 | Bayer Ag | Appareil de séparation pour particules magnétiques en phase liquide |
EP0339980A1 (fr) * | 1988-04-26 | 1989-11-02 | Nippon Telegraph And Telephone Corporation | Microparticules, méthode et appareil pour assembler des spécimens utiles pour étiqueter des réactions immunologiques et méthode et dispositif pour préparer des spécimens |
WO1994019690A1 (fr) * | 1993-02-17 | 1994-09-01 | Cardiovascular Diagnostics, Inc. | Dosage immunologique et dosage par affinite en cascade par voie seche |
EP0687501A2 (fr) * | 1994-06-15 | 1995-12-20 | Precision System Science Co., Ltd. | Méthode de séparation de matière magnétique, utilisant une pipette, et appareils d'analyses cliniques utilisant cette méthode |
Family Cites Families (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CA1110554A (fr) * | 1977-10-25 | 1981-10-13 | John A. Clements | Separateur de phases pour systemes analytiques a debit continu |
EP0030086B2 (fr) * | 1979-11-13 | 1990-03-14 | TECHNICON INSTRUMENTS CORPORATION (a New York corporation) | Ensemble de tubes à essais, kit pour sa fabrication et procédé d'essais immunologiques manuels |
US5076950A (en) * | 1985-12-20 | 1991-12-31 | Syntex (U.S.A.) Inc. | Magnetic composition for particle separation |
DE3764390D1 (de) * | 1986-04-21 | 1990-09-27 | Siemens Ag | Verfahren zur kontinuierlichen separation magnetisierbarer partikel und einrichtung zu seiner durchfuehrung. |
FR2654836B1 (fr) * | 1989-11-17 | 1994-01-28 | Biotrol Sa Laboratoires | Appareil d'execution automatique d'un immunodosage en plusieurs etapes successives d'au moins une substance biologique dans une pluralite d'echantillons biologiques, procede et reactif mettant en óoeuvre ledit appareil. |
US5541072A (en) * | 1994-04-18 | 1996-07-30 | Immunivest Corporation | Method for magnetic separation featuring magnetic particles in a multi-phase system |
US5466574A (en) * | 1991-03-25 | 1995-11-14 | Immunivest Corporation | Apparatus and methods for magnetic separation featuring external magnetic means |
US5186827A (en) * | 1991-03-25 | 1993-02-16 | Immunicon Corporation | Apparatus for magnetic separation featuring external magnetic means |
DE69515675T2 (de) * | 1994-05-11 | 2000-07-20 | Genera Technologies Ltd | Verfahren zum Einfangen von einem Ligand aus einer Flüssigkeit und Vorrichtung zur dessen Ausführung |
US5837144A (en) * | 1994-06-16 | 1998-11-17 | Boehringer Mannheim Gmbh | Method of magnetically separating liquid components |
-
1996
- 1996-05-07 FR FR9605727A patent/FR2748569B1/fr not_active Expired - Fee Related
-
1997
- 1997-05-05 JP JP9539592A patent/JP2000500871A/ja not_active Ceased
- 1997-05-05 EP EP97923133A patent/EP0855030A1/fr not_active Withdrawn
- 1997-05-05 US US08/952,493 patent/US6143577A/en not_active Expired - Fee Related
- 1997-05-05 WO PCT/FR1997/000794 patent/WO1997042503A1/fr not_active Application Discontinuation
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0206077A2 (fr) * | 1985-06-22 | 1986-12-30 | Bayer Ag | Appareil de séparation pour particules magnétiques en phase liquide |
EP0339980A1 (fr) * | 1988-04-26 | 1989-11-02 | Nippon Telegraph And Telephone Corporation | Microparticules, méthode et appareil pour assembler des spécimens utiles pour étiqueter des réactions immunologiques et méthode et dispositif pour préparer des spécimens |
WO1994019690A1 (fr) * | 1993-02-17 | 1994-09-01 | Cardiovascular Diagnostics, Inc. | Dosage immunologique et dosage par affinite en cascade par voie seche |
EP0687501A2 (fr) * | 1994-06-15 | 1995-12-20 | Precision System Science Co., Ltd. | Méthode de séparation de matière magnétique, utilisant une pipette, et appareils d'analyses cliniques utilisant cette méthode |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7651838B2 (en) | 2001-04-30 | 2010-01-26 | Institut National De La Sante Et De La Recherche Medicale | Prenatal diagnosis method on isolated foetal cell of maternal blood |
Also Published As
Publication number | Publication date |
---|---|
EP0855030A1 (fr) | 1998-07-29 |
US6143577A (en) | 2000-11-07 |
FR2748569B1 (fr) | 1998-08-07 |
FR2748569A1 (fr) | 1997-11-14 |
JP2000500871A (ja) | 2000-01-25 |
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