EP1877188A2 - Procede de depot d'echantillons dans des modules et adaptateur associe - Google Patents

Procede de depot d'echantillons dans des modules et adaptateur associe

Info

Publication number
EP1877188A2
EP1877188A2 EP06722826A EP06722826A EP1877188A2 EP 1877188 A2 EP1877188 A2 EP 1877188A2 EP 06722826 A EP06722826 A EP 06722826A EP 06722826 A EP06722826 A EP 06722826A EP 1877188 A2 EP1877188 A2 EP 1877188A2
Authority
EP
European Patent Office
Prior art keywords
adapter
adapter according
module
liquid
sample
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP06722826A
Other languages
German (de)
English (en)
Inventor
Jürgen HUBBUCH
Tim Schroeder
Piere Schulze Wierling
Matthias Bensch
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Forschungszentrum Juelich GmbH
Atoll GmbH
Original Assignee
Forschungszentrum Juelich GmbH
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Forschungszentrum Juelich GmbH filed Critical Forschungszentrum Juelich GmbH
Priority to EP10009331A priority Critical patent/EP2258481A3/fr
Priority to EP10009335A priority patent/EP2266694A3/fr
Publication of EP1877188A2 publication Critical patent/EP1877188A2/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/24—Automatic injection systems
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5025—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures for parallel transport of multiple samples
    • B01L3/50255—Multi-well filtration
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/60—Construction of the column
    • G01N30/6004—Construction of the column end pieces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/026—Fluid interfacing between devices or objects, e.g. connectors, inlet details

Definitions

  • the invention relates to a method for sample application in modules and an adapter.
  • a module In laboratory operation, especially in high-throughput screening (HTS) methods, liquid samples of the prior art are supplied by means of pipetting modules, in particular kits, in which the liquid samples are filtered, adsorbed or eluted, for example.
  • a module is understood to be an open container with an exit and inlet for liquids, for example, but not limited to, a columnar form which is filled with a stationary porous solid phase.
  • a module is to be understood as a kit.
  • the bottom of this vessel is liquid permeable.
  • the solid phase can be designed differently and can be, for example, but not limited to, a filter, a membrane, a liquid-distributing or a chromatographic carrier material.
  • a module is understood to mean a kit, for example, but not limited to the purification of macromolecules (proteins or nucleic acids), filtration of suspensions, cultivation of cells or in medical technology. Applications are found, for example, in the reprocessing of products from biological technological processes, purification of proteins or in medical technology, eg in the analysis of blood samples.
  • the driving force that causes the liquid to flow through the porous solid phase is applied either by centrifugation or by applying a vacuum or overpressure.
  • the overpressure is applied at the top of the module, the vacuum is applied at the bottom of the module.
  • the samples are typically fed into the modules in batch mode.
  • the sample volume that can be loaded into the module is limited by the volume of the modules. If a larger sample volume than the volume of the module to be given, the sample task must be paragraph by paragraph. A portion of the sample volume is dispensed and the sample volume is flowed through the module by applying a driving force. This process is repeated until the entire sample volume has been applied.
  • the standard module used is usually cylindrical or conical vessels with inserts. The latter can be floor inserts such as filters or membranes, but also chromatographic materials. All inserts are commercially available. The invention is particularly directed to the handling of commercial modules, but it should not be limited to their use.
  • pipetting needle is used, but it also includes other substantially needle-shaped constructions, such as pipette tips through which samples can be dispensed.
  • the samples are conveyed by means of pipetting needles in batch mode in the modules.
  • this mode of operation has the following disadvantages:
  • the quality of the tests performed, the time savings and handling and the quality of the samples obtained should be improved.
  • kits / modules it is now possible with the use of kits / modules to enforce larger quantities of samples in a shorter time with an improved product quality.
  • Fig.l A one-piece adapter.
  • Fig.2 A two-piece adapter.
  • Fig.4 A detailed view of the adapter.
  • Fig.5 An exemplary embodiment of the adapter according to the invention.
  • FIG. 1 shows a one-part embodiment of the adapter 1 according to the invention. It has an inlet 2 and an outlet 3.
  • the inlet 2 opens into an upper bore 4, which continues in a lower bore 7, which widens in the form of a funnel 8.
  • a pipetting needle can be received in the bore 4.
  • a thread 9 is optionally provided in the lower part of the adapter 1, a thread 9 is optionally provided in a cavity 5 between the lower and the upper bore 4, 7 there is an O-ring 6, which can receive a pipetting needle and is exemplary for a sealant.
  • FIG. 2 shows a two-part embodiment of the adapter 1 according to the invention with different lower parts IIa, IIb and an upper part 10 which can be inserted into these lower parts IIa, IIb.
  • the upper part 10 has a funnel-shaped inlet 2 for receiving a pipetting needle. It has a region 12 which can be inserted substantially in a fit-fit manner into the region 13 of the lower part which accommodates the upper part 10.
  • the upper part 10 receiving portion 13 of the Lower part IIa, IIb has a constriction 14 which can accommodate an O-ring.
  • the upper part 10 receiving portion 13 of the lower part IIa, IIb narrows in the form of the lower bore 7, which can receive the pipetting needle and through which further flows the exiting liquid.
  • the lower bore 13 expands at the exit in the form of a funnel 8 or in the form of a cylindrical widening 15.
  • Figure 3 shows an embodiment of the adapter 1 according to the invention, which has two inputs 2 for the liquid.
  • the part 16a) shows a side view and the part 16b) is a front view.
  • FIG. 3 a) b) the same reference numerals are assigned to the same device features.
  • Figure 4 shows a concrete embodiment of an inventive adapter 1 in the views a), b), c) and d), wherein a) and b) the lower part and Figure c) and d) is the upper part of the two-part embodiment, and a representation of the matching O-rings in Figures e) and f) and a filter in Figure g).
  • Figure 4b) shows a section of Figure 4a) and Figure 4d) shows a section of Figure 4c).
  • the same device features have the same reference numerals.
  • FIG. 5 shows an adapter according to the invention with a pipetting needle 18, which is inserted into the upper part 10 of a two-part adapter and whose tip projects into the lower part 11 of the two-part adapter 1.
  • an O-ring 6 In the space between the upper and lower parts 10, 11 of the adapter there is an O-ring 6.
  • the adapter 1 is on a module / kit 20 plugged and sealed with respect to the entrance area of the module / kit 20 with another O-ring.
  • a liquid sample is usually introduced through a pipetting needle or a pipette tip through an adapter 1 in the module / kit 20.
  • a module 20 according to the invention is an open container with liquid inlet and outlet, by way of example but not limitation, to be understood in column form, which is filled with a stationary porous solid phase.
  • the bottom of this vessel is liquid permeable.
  • the solid phase can be designed differently and can be, for example, but not limited to, a filter, a membrane, a liquid-distributing or a chromatographic carrier material.
  • kits may be taken to mean an analysis kit, such as, but not limited to, a diagnostic kit, an RNA isolation kit, or a PCR kit or kits for the separation or purification of proteins.
  • Such kits can be used in the field of microbiology, biotechnology or medical technology.
  • the module according to the invention therefore has, in a preferred embodiment, essentially the dimensions customary for kits. Typical target applications of kits are rapid separation, good separation efficiency, and high recovery of valuable samples.
  • the typical formats of kits are one-of-a-kind, 8-strip, 9-bit Mirotiter (8x12) kits.
  • kits for example, in the selective binding (adsorption) of certain proteins on a mostly solid phase (also called adsorbent), z. B. membrane or chromatography adsorber particles.
  • adsorbent can selectively adsorb certain proteins, all other samples flow through the kit without binding.
  • the bound proteins can be dissolved, eluted or desorbed and are then in a better purity.
  • the product may represent the molecules flowing through while unwanted components bind to the solid phase.
  • pipetting needle In the following the term pipetting needle is used. However, it also includes other essentially needle-shaped constructions, such as pipette tips, through which samples can be dispensed.
  • the adapter 1 establishes a liquid-tight connection between the pipetting needle and the module 20, through which the input of the sample can take place.
  • the adapter 1 is integrally formed in one embodiment and includes an input 2 and an output third
  • the output 3 is liquid-tightly connected to the module 20.
  • the outlet 3 or the lower part of the adapter 1 can be formed so as to be insertable and liquid-tight in the module 20.
  • at least one lower portion of the adapter 1 may have a thread 9, which may be screwed into the entrance of a module 20.
  • the thread 9 is exemplary of accurately sealing connections.
  • the outlet 3 of the adapter 1 is preferably provided with sealing means for liquid-tight connection to the module 20.
  • sealing means for liquid-tight connection to the module 20.
  • These can be, for example, an O-ring, adhesive films, Be foils or a diaphragm. Also conceivable is an external thread, which is possibly surrounded by a sealant.
  • the adapter 1 can be a firmly seated integral component of the module or kit, so that it is inseparably connected. This applies to both the one-part and the two-part embodiment.
  • the outlet 3 preferably has a sample outlet which assumes a geometry which allows a uniform liquid outlet over the entire cross section of the sample outlet.
  • This liquid outlet is preferably uniform over the entire entrance surface of the module / kit 20.
  • This can be effected, for example, in that the sample outlet forms a substantially conically expiring cavity, for example in the form of a funnel 8 or a step-shaped cavity 15, which tapers inwardly substantially radially symmetrically from the outlet end.
  • Alternative options for uniform liquid distribution over the outlet cross section may be inserts, such as membranes, perforated plates or filter discs. These are exemplary of means for uniform liquid distribution over the cross section of the output.
  • the adapter 1 is designed such that the sample outlet or the outlet 3 with the bottom part of the module 20, for example the filter, the membrane or the chromatography carrier substance does not allow a dead volume or a small dead volume.
  • the sample outlet or the outlet 3 is substantially flush with the bottom of the module, or that only a small Liquid volume, preferably without gas phase, located above the bottom of the module.
  • the outer diameter of the outlet 3 corresponds essentially to the inner diameter of the module 20 and is therefore preferably oriented to the dimensions of commercially available and normalized inner diameters of the kits. These are for example 4 mm, 6 mm or 6.5 mm. But it can also be freely selectable outer diameter can be realized, which correspond to the inner diameters, for example, self-made and non-normalized vessels. Overall, it depends on the function of the liquid-tight contact between the adapter 1 and the module 20.
  • the inlet 2 of the adapter 1 is designed so that it allows a liquid-tight recording of the sample.
  • the inner diameter of the input region of the adapter 1 can substantially correspond to the outer diameter of the pipetting needle, so that ideally a pass-fit connection is established.
  • the adapter 1 may be sealed liquid-tight at its entrance 2 with a septum.
  • the bore 4 may have a larger inner diameter than the bore. 7
  • an O-ring is received in a constriction 14, which surrounds the pipetting needle.
  • the inner diameter of the O-ring is smaller than the outer diameter of the pipetting del, so that the O-ring covers the pipetting needle and seals, so that the liquid can flow only in the flow direction.
  • the input 2 of the adapter 1 can be provided with sealing means which the liquid-tight input of
  • the inlet 2 may be provided with a liquid-tight connection, which allows the attachment of a tubular sample supply line.
  • This sample supply line can be a flexible, preferably pressure-resistant line which, for example, allows connection to a pump system for conveying liquids or to other method steps upstream of the module / kit 20.
  • Such lines can be fixed, for example, by crimping screwed connections with union nuts and possibly additional sealing means.
  • the adapter is sealed against the module 20 and on the other hand the pipetting needle is sealed against the adapter.
  • the adapter 1 connected to the module 20 preferably allows an internal pressure load of 0.2 MPa, better up to 0.3 or 0.5 MPa, most preferably 1 MPa.
  • the adapter 1 is formed in two parts and comprises a lower, the module 20 facing part and an upper separate input part. In this case, it is possible to combine various input and output parts with each other, so that the adapter 1 can be used in a variety of ways.
  • the upper and lower parts 10, 11 of the two-part adapter 1 can be connected to one another, for example, by a screw connection or a plug connection.
  • This compound is in turn preferably formed liquid-tight and preferably comprises sealing means. It is preferably pressure-resistant in the same way as the one-piece mold.
  • the inlet 2 of the adapter 1 can be designed such that it can accommodate at least two pipetting needles or supply lines.
  • an adapter can have 1 n inputs, which open into a mixing channel 17, so that mixing of different components is possible.
  • n may be 10-2, preferably 5-2, more preferably 2. This allows the mixing of different sample liquids together. The supply of these liquids can follow certain metering rules, so that defined compositions can be achieved.
  • At least two adapters 1 can be arranged together, so that a plurality of modules 20, which are arranged in rows or a preferably rectangular grid, can be operated simultaneously.
  • This arrangement may include 2, 4, 8, 96 or more adapters 1 in a row or other arrangement, such as a circle or circle segment or other rounding.
  • an adapter in this case, both an adap- ter as well as the arrangement by means of a mixing channel 17 to understand.
  • Preferred is an arrangement in rows or rectangular structures.
  • the adapter according to the invention makes possible a continuous operation in that the sample is introduced into the kit or module 20, preferably under pressure, in a continuous process. This eliminates the need for batch operation in which the module 20 is either emptied repeatedly by aspiration or pressurization applied after removal of the pipette. It can also be dispensed with a centrifugation. All these process steps involve time-consuming and material-intensive or machine-consuming steps which can be saved. Fractionation of discontinued samples can be omitted.
  • the invention also encompasses a method for filling modules 20, in which the samples metered into the modules 20 are filled into the modules in flow mode according to the invention.
  • the method is characterized in particular by the samples being introduced into the module 20 continuously and in a liquid-tight manner by means of a pipetting needle.
  • the sample is conveyed into the module 20 during continuous operation.
  • the sample is in this case conveyed by means of a pump through a pipetting needle via a liquid-tight connection in the module 20 and passes through this.
  • the bottom of the module 20 may be formed as a filter, membrane or Adsor- bens.
  • the sample then leaves the module 20 and can be fed to further process steps.
  • the sample is passed over an adapter according to the invention.
  • various samples may be fed in parallel to a module 20 in which they mix.
  • a particular advantage is the quality improvement of the examination or the procedure.
  • This quality improvement is z. B. by the interference-free passage of the sample through the medium, for example, the filter, the chrorata-tographischen solid or the membrane causes, as an uninterrupted liquid flow leads to the best separation or elution results.
  • the trouble-free course is an important measure for quality assurance.
  • Precise flow parameters can be set, for example by conveying a delivery pump according to requirements. For the first time, it is possible for the loading of kits or modules 20 to mix the sample, which is made possible by setting elution conditions in gradients or step form. Furthermore, samples or substances reacting with one another can be mixed.
  • samples can be placed in kits or modules 20.
  • the applications are in particular characterization and development of modules / kits 20, examination of the methods represented by these modules / kits 20 or the application of these modules / kits 20 for treating or cleaning samples.
  • an adapter has been constructed which allows directly small columns to be used with commercially available formats on pipetting tips used on HTS platforms or approach columnar containers and operate them in flow mode.
  • the adapter can be used as a single piece, strip (linear combination of adapters) or in multiwell format.
  • the adapter offers the possibility of connecting it to all conventional formats of multi-well plates, columns, membrane stacks or containers, while it can be approached by pipetting tips at the upper end by means of a sealing device.
  • the liquid distribution at the lower end is by conventional techniques such.
  • the upper adapter part offers the possibility to connect common connections of chromatography systems and to operate the respective application with them.
  • the adapter allows the control of miniaturized chromatography columns to operate under flow. This allows a parallelization of chromatographic experiments, which can be carried out on pipetting platforms (HTS applications) analogous to all common techniques on a laboratory scale. This technique gives better results in the purification of molecules on a micro scale, since the separation potential of columns can be exploited. Furthermore, this technique allows a parallelized use of columns for the development of chromatography processes. The technique also allows different chromatographic materials to be tested for their quality for various applications.
  • Another example is found in the continuous control of membranes or membrane stacks.
  • the application bandwidth is similar to that described above.
  • the technique also allows to test various membrane materials for their quality for different applications.
  • Filter plates can be loaded continuously by this method and so be applied to larger amounts of material to be filtered. This is suitable both for a larger amount of filtrate and retentate (eg cells, inclusion to generate bodies).
  • the technique makes it possible to continue to test ver ⁇ different filter materials for quality for Various ⁇ ne applications.

Abstract

Procédé de dépôt d'échantillons dans des modules, en particulier dans des kits, et adaptateur associé. L'adaptateur (1) selon la présente invention est ainsi conçu qu'il permet la réception étanche aux liquides d'une aiguille d'injection. Il est relié de manière étanche aux liquides avec le module ou kit (20) se trouvant en dessous et permet un écoulement continu d'échantillons.
EP06722826A 2005-05-03 2006-04-25 Procede de depot d'echantillons dans des modules et adaptateur associe Withdrawn EP1877188A2 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP10009331A EP2258481A3 (fr) 2005-05-03 2006-04-25 Adaptateur
EP10009335A EP2266694A3 (fr) 2005-05-03 2006-04-25 Adaptateur

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
DE102005020983 2005-05-03
DE102005026585A DE102005026585A1 (de) 2005-05-03 2005-06-08 Verfahren zur Probenaufgabe in Module sowie Adapter
PCT/DE2006/000708 WO2006116964A2 (fr) 2005-05-03 2006-04-25 Procede de depot d'echantillons dans des modules et adaptateur associe

Publications (1)

Publication Number Publication Date
EP1877188A2 true EP1877188A2 (fr) 2008-01-16

Family

ID=36691335

Family Applications (3)

Application Number Title Priority Date Filing Date
EP06722826A Withdrawn EP1877188A2 (fr) 2005-05-03 2006-04-25 Procede de depot d'echantillons dans des modules et adaptateur associe
EP10009331A Withdrawn EP2258481A3 (fr) 2005-05-03 2006-04-25 Adaptateur
EP10009335A Withdrawn EP2266694A3 (fr) 2005-05-03 2006-04-25 Adaptateur

Family Applications After (2)

Application Number Title Priority Date Filing Date
EP10009331A Withdrawn EP2258481A3 (fr) 2005-05-03 2006-04-25 Adaptateur
EP10009335A Withdrawn EP2266694A3 (fr) 2005-05-03 2006-04-25 Adaptateur

Country Status (4)

Country Link
US (1) US8474336B2 (fr)
EP (3) EP1877188A2 (fr)
DE (1) DE102005026585A1 (fr)
WO (1) WO2006116964A2 (fr)

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US11731062B2 (en) 2021-01-20 2023-08-22 Asahi Kasei Bioprocess America, Inc. Components that facilitate maintenance of chromatography and synthesis columns

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Also Published As

Publication number Publication date
DE102005026585A1 (de) 2006-11-09
EP2258481A3 (fr) 2011-12-07
EP2266694A2 (fr) 2010-12-29
EP2266694A3 (fr) 2011-12-07
US8474336B2 (en) 2013-07-02
US20100242634A1 (en) 2010-09-30
WO2006116964A2 (fr) 2006-11-09
EP2258481A2 (fr) 2010-12-08
WO2006116964A3 (fr) 2007-02-15

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