U.S. patent application number 10/491025 was filed with the patent office on 2005-04-07 for electrolyser.
Invention is credited to Benjamin Ross, Herbert, Matthew charles Archer, Durack, Rummery, Gerard Michael, William Samuel, Hunter.
Application Number | 20050072666 10/491025 |
Document ID | / |
Family ID | 3831839 |
Filed Date | 2005-04-07 |
United States Patent
Application |
20050072666 |
Kind Code |
A1 |
Rummery, Gerard Michael ; et
al. |
April 7, 2005 |
Electrolyser
Abstract
An electrolyser for separating a mixture of components using
isoelectric focusing in a liquid media comprises a series of blocks
defining chambers. The chambers are separable by isoelectric
membranes of known pH. End chambers are located at each end of the
series of chambers. The blocks are clamped together using four tie
rods to make the apparatus substantially liquid tight. An
electrodes is provided in each end chamber for use in applying an
electric field across the series of chambers. One of the tie rods
is conducting and carries current from an electrode at one end of
the apparatus to the opposite end of the apparatus. A lever
operated clamping mechanism is provided for tensioning the tie rods
and clamping the chambers together between the end blocks in a
liquid tight fashion.
Inventors: |
Rummery, Gerard Michael;
(Queanbeyan NSW, AU) ; William Samuel, Hunter;
(Jan Juc Vic, AU) ; Benjamin Ross, Herbert; (North
Epping, AU) ; Matthew charles Archer, Durack; (Glebe,
AU) |
Correspondence
Address: |
BOZICEVIC, FIELD & FRANCIS LLP
1900 UNIVERSITY AVENUE
SUITE 200
EAST PALO ALTO
CA
94303
US
|
Family ID: |
3831839 |
Appl. No.: |
10/491025 |
Filed: |
November 22, 2004 |
PCT Filed: |
September 30, 2002 |
PCT NO: |
PCT/AU02/01316 |
Current U.S.
Class: |
204/244 ;
204/253 |
Current CPC
Class: |
B01D 57/02 20130101;
G01N 27/44795 20130101 |
Class at
Publication: |
204/244 ;
204/253 |
International
Class: |
C25B 009/00 |
Foreign Application Data
Date |
Code |
Application Number |
Sep 28, 2001 |
AU |
PR 8009 |
Claims
1. An apparatus for separating a mixture of components using
isoelectric focusing in a liquid media, the apparatus comprising: a
series of blocks defining chambers, the chambers being separable by
isoelectric membranes of known pH; end chambers located at each end
of the series of chambers; at least two tie rods for use in
clamping the blocks together to make the apparatus substantially
liquid tight; and electrodes for use in applying an electric field
across the series of chambers; wherein at least one of the tie rods
is conducting and carries current from an electrode at one end of
the apparatus to the opposite end of the apparatus.
2. The apparatus of claim 1 wherein the conducting tie rod is a
metal rod which is in an insulating material along a substantial
portion of its length.
3. The apparatus of claim 2 wherein the conducting tie rod is a
steel rod.
4. The apparatus of claim 1 including four tie rods.
5. An apparatus for separating a mixture of components using
isoelectric focusing in a liquid media, the apparatus comprising: a
series of blocks defining chambers, the chambers being separable by
membranes and capable of being clamped together in a watertight
arrangement, the apparatus including end blocks between which the
series of chambers may be held by means of a plurality of tie rods
extending through the blocks, wherein a lever operated clamping
mechanism is provided for tensioning the tie rods and clamping the
chambers together between the end blocks in a liquid tight
fashion.
6. The apparatus of claim 1 wherein a lever operated clamping
mechanism is provided for tensioning the tie rods and clamping the
chambers together between the end blocks in a liquid tight
fashion.
7. The apparatus of claim 5 wherein one end of each of the tie rods
is fixed with respect to one end block of the apparatus.
8. The apparatus of claim 7 wherein the other end of each of the
tie rods defines enlarged end portions and the other end block of
the apparatus comprises a housing and a plate moveable relative to
the housing and wherein the plate defines a series of apertures
adapted to receive the ends of the tie rods and engage with the
enlarged portions of the tie rods and wherein the clamping means is
arranged to move the plate away from the housing using a cam
mechanism or the like to tension the rods.
9. The apparatus of claim 8 wherein the apertures are generally
keyhole shaped.
10. The apparatus of claim 9 wherein the engagement of the ends of
the tie rod to the plate is by means of a twist fitting with the
tie rods being inserted with the plate being rotationally off-set
relative to the apparatus and rotatable to align the cross-section
of the plate with the blocks and engage the enlarged portions of
the tie rods behind narrower portions of the keyhole shaped
apertures.
11. An apparatus for separating a mixture of components using
isoelectric focusing in a liquid media, the apparatus comprising a
series of blocks defining chambers, the chambers being open on
front and rear faces of the blocks, the chambers being separable by
membranes disposed between adjacent blocks, the blocks being
capable of being clamped together in a watertight arrangement,
wherein the membranes are retained in washers each of which defines
a recess for receiving a membrane and locating the same in correct
alignment with the chambers.
12. The apparatus of claim 11 wherein one face of each block
defines a recess into which a corresponding annular protrusion
locates and wherein an end wall of the recess defines a stepped
portion and a front face of the annular protrusion defines a mating
stepped portion the washer being stepped also and locating in use
between the end wall and the front face of the annular
protrusion
13. A dummy block for use with the apparatus of claim 1, the block
defining a pair of spaced apart electrical contacts which are
biased apart but electrically connected which is adapted to replace
one of the blocks in the apparatus while still allowing the passage
of current along the chambers of the apparatus, in use.
14. An apparatus for separating a mixture of components using
isoelectric focusing in a liquid media, the apparatus comprising: a
series of blocks defining chambers, the chambers being separable by
isoelectric membranes of known pH; end chambers located at each end
of the series of chambers with each chamber defining a well opening
into the base of the chamber, in use; at least two tie rods for use
in clamping the blocks together to make the apparatus substantially
liquid tight; electrodes for use in applying an electric field
across the series of chambers; and a power supply defining a
platform on which the apparatus may be placed in use, the power
supply is arranged to supply power to the electrodes; wherein a
magnetic plate is located in the well wherein movement of the plate
is accomplished by a solenoid disposed under a floor of the
platform and means for periodically reversing the polarity of the
solenoid.
15. The apparatus of claim 14 wherein the diameter of the well is
slightly larger than the diameter of the magnetic plate such that
the magnetic plate is prevented from turning over but is free to
move along the central axis of the well.
16. The apparatus of claim 14 wherein the well defines an internal
flange where it meets the base of the chamber dimensioned to
prevent egress of the plate from the well.
17. A method of agitating fluid contained in chambers of the
apparatus of claim 1 comprising the steps of providing a well
beneath and open to a chamber in the block; providing a stirring
means in the form of a ferro-magnetic disk in the well; providing a
solenoid disposed beneath the well and magnetic disk in operation;
and periodically reversing the polarity of the solenoid thereby
causing movement of the magnetic plate.
Description
FIELD OF THE INVENTION
[0001] The present invention relates to a improved apparatus for
electrophoresis and in particular to an apparatus of the type
commonly referred to as an "electrolyser".
BACKGROUND OF THE INVENTION
[0002] International patent application No PCT/AU00/01391 filed in
the name of the applicant of the present invention relates to such
an electrolyser and to a method of using that electrolyser for
sub-fractionation and subsequent separation of fractions from
highly complex protein/peptide mixtures, such as those found in
total cell lysates, body fluids and tissue extracts in general.
[0003] That application discloses an electrolyser in the form of a
series of chambers separated by isoelectric membranes with end
chambers located either end of the series of chambers. Electrode
means for applying an electric field across the series of chambers
are provided. A means for agitating any liquid media within the
chambers without recirculation of the liquid is also disclosed. The
means for agitating is typically a magnetic stirrer.
[0004] The present invention is directed to improvements to the
apparatus of the type disclosed in PCT/AU00/01391, the entire
contents of which are incorporated herein by reference.
[0005] Any discussion of documents, acts, materials, devices,
articles or the like which has been included in the present
specification is solely for the purpose of providing a context for
the present invention. It is not to be taken as an admission that
any or all of these matters form part of the prior art base or were
common general knowledge in the field relevant to the present
invention as it existed in Australia before the priority date of
each claim of this application.
SUMMARY OF THE INVENTION
[0006] In a first aspect, the present invention provides an
apparatus for separating a mixture of components using isoelectric
focusing in a liquid media, the apparatus comprising:
[0007] a series of blocks defining chambers, the chambers being
separable by isoelectric membranes of known pH;
[0008] end chambers located at each end of the series of
chambers;
[0009] at least two tie rods for use in clamping the blocks
together to make the apparatus substantially liquid tight; and
[0010] electrodes for use in applying an electric field across the
series of chambers:
[0011] characterised in that at least one of the tie rods is
conducting and carries current from an electrode at one end of the
apparatus to the opposite end of the apparatus.
[0012] Using one of the tie rods to carry current, allows power
supply sockets for supplying power to the electrodes to be located
at the same end of the apparatus.
[0013] This has the advantage that the apparatus is easier to use.
By providing all the electrical connections on one end of the
apparatus, the apparatus becomes suitable for use on a multi-use
platform power supply and cooling system which provides electrical
contacts along one side only of the platform.
[0014] The conducting tie rod will preferably be a steel rod, and
is most preferably covered in an insulator along a substantial
portion of its length.
[0015] It is preferred that there are four tie rods, although only
one of the tie rods will typically be used to conduct
electricity.
[0016] In a second aspect of the present invention, there is
provided an apparatus for separating a mixture of components using
isoelectric focusing in a liquid media comprising a series of
blocks defining chambers, the chambers being separable by membranes
and capable of being clamped together in a watertight arrangement,
the apparatus including end blocks between which the series of
chambers may be held by means of a plurality of tie rods extending
through the blocks characterised in that a lever operated clamping
mechanism is provided for tensioning the tie rods and clamping the
chambers together between the end blocks in a liquid type
fashion.
[0017] In a particularly preferred embodiment, one end of each of
the tie rods is fixed with respect to one end block of the
apparatus. The other end of each of the tie rods defines enlarged
end portions. The other end block of the apparatus comprises a
housing and a plate moveable relative to the housing. The plate
defines a series of keyhole shaped free apertures adapted to
receive the ends of the tie rods and engage with the enlarged
portions of the tie rods. The damping means is arranged to move the
plate relative to the housing using a cam mechanism or the
like.
[0018] In a particularly preferred embodiment, in cross-section the
blocks and the plate are generally rectangular, typically square,
in lateral cross-section. The engagement of the ends of the tie rod
to the plate is by means of a twist fitting with the tie rods being
inserted with the plate being rotationally off-set relative to the
apparatus and rotatable to align the cross-section of the plate
with the blocks and engage the enlarged portions behind narrower
portions of the keyhole shaped apertures.
[0019] The use of a lever makes the assembly of the apparatus much
quicker than the traditional method which relies on wing nuts to
tension the tie rods and also allows an increased pressure to be
used and thus improves the sealing of the apparatus.
[0020] In a yet further aspect, the invention provides an apparatus
for separating a mixture of components using isoelectric focusing
in a liquid media, the apparatus comprising a series of blocks
defining chambers, the chambers being open on front and rear faces
of the blocks, the chambers being separable by membranes disposed
between adjacent blocks, the blocks being capable of being clamped
together in a watertight arrangement, characterised in that the
membranes are retained in washers each of which defines a recess
for receiving a membrane and locating the same in correct alignment
with the chambers.
[0021] Typically, one face of each block defines a recess into
which a corresponding annular protrusion locates and wherein an end
wall of the recess defines a stepped portion and a front face of
the annular protrusion defines a mating stepped portion the washer
being stepped also and locating in use between the end wall and the
front face of the annular protrusion.
[0022] The apparatus is typically designed to receive a set number
of chamber blocks, typically seven, however, in some cases fewer
chambers will be required.
[0023] Therefore, in related aspect, the present invention provides
a dummy block defining a pair of spaced apart electrical contacts
which are biased apart but electrically connected which is adapted
to replace one of the chamber blocks in the apparatus while still
allowing the passage of current along the chambers of the
apparatus, in use.
[0024] The invention also provides a novel method of agitating
fluid contained in chambers of the apparatus. In this aspect of the
present invention, a well is provided beneath and open to the
chamber in the block. The stirring means is a magnetic disk and
means for agitating the stirring means comprise a solenoid disposed
beneath the magnetic disk, in operation. Typically this solenoid
will be located in the platform which supplies any necessary
cooling and power to the apparatus during use. The polarity of the
solenoid is periodically reversed thereby causing movement of the
magnetic plate.
[0025] Whilst a single solenoid may perform this function, a
plurality of solenoids could be used.
[0026] In a particularly preferred embodiment, the magnetic plate
is encased in a housing which is marginally smaller than the well
in which the plate is located such that the magnetic plate is
prevented from turning over but is free to move along the
(vertical) axis of the well. Preferably, the top of the well is
partly closed to prevent the magnetic plate from leaving the
well.
BRIEF DESCRIPTION OF THE DRAWINGS
[0027] Specific embodiments of the invention will now be described,
by way of the example only, and with reference to the accompanying
drawings in which:
[0028] FIG. 1 is a perspective view of an apparatus embodying the
present invention;
[0029] FIG. 2 is a perspective view of a chamber block which forms
part of the apparatus shown in FIG. 1;
[0030] FIG. 3 is an underneath plan view of two chamber blocks
engaged together;
[0031] FIG. 4 is a section through lines IV-IV of FIG. 3;
[0032] FIGS. 5a, 5b, and 5c are a perspective view, front elevation
and a section through a stepped washer which forms part of the
apparatus of FIG. 1.
[0033] FIG. 6 is a perspective view of parts of an apparatus
embodying the present invention illustrating internal features of
one end block in particular;
[0034] FIG. 7 is an end view of the apparatus shown in FIG. 1 with
part of the housing of the end block of the apparatus removed;
[0035] FIG. 8 is an end view of a plate;
[0036] FIG. 9 is a perspective view of an end plate illustrating
connectors for a conducting tie rod;
[0037] FIG. 10 is a perspective view of a dummy block;
[0038] FIG. 11 is a front view of the dummy block of FIG. 10;
[0039] FIG. 12 is a cross section on lines XII-XII of FIG. 11;
[0040] FIG. 13 shows the apparatus of FIG. 1 located on a platform
with a top plate of the platform removed; and
[0041] FIG. 14 is a top plan view of the platform of FIG. 13;
[0042] FIG. 15 is a top plan view of the apparatus of FIG. 1
located on a complete platform;
[0043] FIG. 16 is a cross sectional view on XVI-XVI of FIG. 15;
[0044] FIG. 17 is an enlarged view of part of FIG. 16; and
[0045] FIG. 18 illustrates an alternative stirring mechanism for
the apparatus.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT
[0046] Referring to the drawings, FIG. 1 shows an assembled
multi-compartment electrolyser apparatus generally indicated at 10.
The apparatus includes seven identical chamber blocks 12, clamped
between end blocks 14, 16.
[0047] FIGS. 2 to 4 shows the chamber blocks 12 in more detail.
Each chamber block is generally square in lateral/transverse cross
section and is generally hollow so that the central area of the
block defines a chamber 18 As is best seen in FIG. 3 an annular
ring 20 projects from one (front) end face 22 of the block. The
opposite (rear) end face 24 defines a corresponding recess adapted
to receive the projecting ring of an adjacent block. A through bore
is defined when a plurality of blocks are engaged end to end as
illustrated in FIG. 3. Four narrower through bores 25 are defined
near the corners of the end faces of each blocks. These bores 25
receive tie rods 40 best seen in FIGS. 6 and 7 which are used to
align the bores together and link and compress the chambers
together.
[0048] The front face 22 of each chamber block also defines two
additional cylindrical protrusions 28 which are received in
corresponding recesses located on the rear face of a contiguous
block for ensuring accurate alignment of the blocks.
[0049] At the top 30 of each block there is a sample inlet in the
form of an aperture 32 for sample entry into the chamber 18. The
top of each block is closed with a lid 34. Each chamber also
defines a generally cylindrical shallow well or recess 36 which is
typically coaxial with the sample inlet 32. In use, the well
receives a magnetic disk which functions as a stirrer when placed
in a magnetic field generated by a solenoid whose polarity is
periodically reversed. alternatively a standard bipolar magnetic
stirrer may be used.
[0050] The cylindrical recess defined in the rear face is adapted
to receive a washer and membrane assembly 38. As is best seen in
FIG. 5 the washer and membrane assembly comprises two stepped
washers, which may be made of rubber or another suitable material
and a circular membrane or septum 39 located and held between the
two washers and which in use closes one end of the chamber 18. The
fact that the washers are stepped holds the septa in position away
from the walls of the chamber/block, improves sealing and makes
assembly of the apparatus easier. It will be readily appreciated
that when two blocks are juxtaposed as shown in FIG. 4, the
membranes will function as dividing walls and will separate one
chamber 18 from the chamber 18 of the adjacent block. The septa
between the various chambers are isoelectric, buffering membranes,
cast onto a supportive glass fibres or other suitable material. An
O-ring may be disposed outside the membrane/washers to provide
additional sealing. Such membranes are flow tight and ensure proper
pH control.
[0051] The multi-compartment electrolyser apparatus is assembled by
placing septa/dividing walls between adjacent chambers, inserting a
tie rod through the aligned holes in each chamber block and
compressing the chamber blocks together between end blocks 14, 16
in a manner which will be described in more detail below.
[0052] The clamping mechanism will now be described in more detail.
One end of each of the four tie rods 40 is embedded in one of the
end blocks 16 of the apparatus, spaced apart and in the correct
orientation for simultaneous insertion through the bores 25 of one
of the chamber blocks 12.
[0053] The other end block 14 of the apparatus is best described
with reference to FIGS. 7 to 9. It includes a generally square
plate 42 which is movable relative to the housing 44 of the end
block which housing in use, engages against an adjacent cylinder
block 12. For clarity, FIG. 7 shows part of the housing removed.
The end block 42 defines a keyhole shaped aperture 43 in each
corner of the block so that there are four apertures in total. The
keyhole shaped apertures have a relatively wider part 46 and a
relatively narrower part 48 (refer to FIG. 8). Each of the tie rods
has an enlarged portion 50 at the end distal from block 16. The
enlarged portion 50 is small enough to pass through the wider part
46 of the keyhole apertures 43 but is too large to pass through the
narrower part 48 of those keyhole apertures 43. The keyhole
apertures are aligned such that the narrower parts 48 of the
keyhole apertures are located on diagonals "D" passing through the
corners of the plate and the wider portion of the keyhole are all
rotated through an angle ".alpha." relative to the diagonals (refer
to FIG. 8). This enables a bayonet type latching of the plate to
the tie rods in which the plate 42 is first aligned such that it is
off set by angle .alpha. relative to the cross section of the
chamber blocks 14 to allow the tie rods to pass through the keyhole
apertures and then turned through an angle .alpha. so that the
orientation of the square plate 42 is aligned with that of the
chamber blocks 12. In that orientation, the tie rods are locked
behind the plate as the enlarged portions 50 cannot pass through
the narrower part 48 of the keyhole apertures. A lever 60 is linked
to the plate 42 by means of a metal rod and a camming means or
other suitable mechanism is provided such that releasing the lever
allows the plate to move generally towards the chamber blocks 12
whereas closing the lever into the position shown in FIG. 7 tends
to pull the plate away from the chamber blocks 12 thus tensioning
the tie rods and compressing the chambers between the end block 16
and the housing 44 of end block 14.
[0054] FIG. 6 illustrates a flexible springy connector 70 which is
in electrical contact with one of the tie rods 40A which is a
conducting electrode. The connector is connected to a positive
terminal of a power supply and in use current is passed down the
tie rod to a positive electrode (not shown). FIG. 9 shows the
connector 72 for the negative electrode (not shown) located in the
end block 14. This allows both positive and negative electrical
connections to be at the one end of the electrolyser in one socket
76--see FIG. 1.
[0055] A further novel feature of the invention is the use of dummy
blocks 80 illustrated in FIGS. 10 to 12. The dummy blocks are
substantially the same shape and size as the chamber blocks and
define four bores 25 for receiving the tie rods. However, instead
of defining a chamber, the dummy blocks define a contact plate 82
and a contact pin 84 which are biased apart by means of an
electrically conducting spring means 86. If electrophoretic
separation is to be carried out using fewer than seven chambers,
one dummy block is inserted in the apparatus to replace each of the
"missing" chamber blocks.
[0056] FIGS. 13 to 17 illustrate the apparatus of the present
invention disposed on a power supply/platform 100. FIG. 13 shows
the top plate 102 of the platform removed so that magnets 106
mounted on belt 107 driven rotatable turntables 104 can be seen. In
use, as the magnets rotate, bipolar magnetic stirrers located in
the wells of the chambers of the blocks of the electrolyser rotate
to stir the contents of the chamber. The platform 100 includes an
aluminium top plate 102, a Peltier 108 for cooling the plate and a
fan (not shown) for cooling the Peltier.
[0057] The invention provides an alternative novel stirring
mechanism which is illustrated in FIG. 18. A solenoid 120 is
disposed underneath the well 36 of a block which as can be seen in
the Figure, has a top entrance 36a which is narrower than the bore
of the well 36. Inside the well, is located a stirrer means 130
comprising a magnetic plate 132 which is encased in a plastic
coating 134 or the like which defines an annular cylindrical
portion 136 around its extreme and which prevents the stirrer from
over-turning in the well. Periodic reversal of the polarity of the
solenoid 120 causes the stirrer means 130 to move up and down
inside the well and therefore stirs the contents of the
chamber.
[0058] It will be appreciated by persons skilled in the art that
numerous variations and/or modifications may be made to the
invention as shown in the specific embodiments without departing
from the spirit or scope of the invention as broadly described. The
present embodiments are, therefore, to be considered in all
respects as illustrative and not restrictive.
* * * * *