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  Small separation apparatusUSPTO Application #: 20060037860Title: Small separation apparatus Abstract: An apparatus for processing compounds in small volumes by electrophoresis, the apparatus comprising: (a) a cathode in a static cathode buffer zone or compartment; (b) an anode in a static anode buffer zone or compartment, the anode disposed relative to the cathode so as to be adapted to generate an electric field in an electric field area therebetween upon application of a voltage potential between the cathode and anode; (c) a first separation barrier disposed in the electric field area; (d) a second separation barrier disposed between a selected one of the cathode buffer zone and the anode buffer zone and the first barrier so as to define a first interstitial volume or chamber therebetween; wherein in use, electrophoretic buffer is disposed in the cathode buffer zone and the anode buffer zone, a sample constituent is provided to the first interstitial volume; wherein upon application of the voltage potential, a selected separation product is removed from the sample constituent through a selected one of the first and second separation barriers, and provided to a selected one of the cathode buffer and anode buffer zones; and wherein there is substantially no circulation of buffer or sample constituent in the buffer zones or the first interstitial volume. (end of abstract) Agent: Morgan Lewis & Bockius LLP - Washington, DC, US Inventors: David Ogle, Dennis Rylatt USPTO Applicaton #: 20060037860 - Class: 204450000 (USPTO) Related Patent Categories: Chemistry: Electrical And Wave Energy, Non-distilling Bottoms Treatment, Electrophoresis Or Electro-osmosis Processes And Electrolyte Compositions Therefor When Not Provided For Elsewhere The Patent Description & Claims data below is from USPTO Patent Application 20060037860. Brief Patent Description - Full Patent Description - Patent Application Claims
BACKGROUND OF THE INVENTION [0001] This invention relates to an electrophoretic separation apparatus and in particular, to apparatus for treatment or separation of compounds including macromolecules in small volumes. [0002] In the past, the separation of macromolecular solutes was performed by a process known as electrophoretic separation or electrophoresis, in particular, fixed boundary electrophoretic separation. In fixed boundary electrophoresis, a semi-permeable membrane (hereinafter referred to as a separation membrane), acts to separate two streams of liquid carrying macromolecular solutes such as proteins, known as the sample and the downstream. The streams flow between charged electrodes and at least one macromolecular solute migrates across the membrane from one stream to the other stream under the influence of the electric field. The apparatus also includes flow paths for buffer solutions and further semi-permeable membranes, hereinafter referred to as restriction membranes, disposed either side of the separation membrane between the electrodes and the separation membrane to separate the buffer flow paths from the sample and downstream. The restriction membranes allow the passage of ions but not of the relatively larger macromolecules. [0003] Attempts have been made to improve upon such fixed boundary electrophoresis technology. In particular, one such improvement provided a system in which the separation and restriction membranes and sample and separation are contained in a removable and replaceable cartridge. Although this technology provides a substantial improvement, electrophoresis cannot satisfactorily be used for separating very small samples. Even an apparatus of reduced size is unable to separate very small samples, with the smallest practicable sample size being around 6 mL. Therefore, for smaller samples, other separation methods such as chromatography or gel electrophoresis, have to be used. However, such other methods are time consuming. For example, in gel electrophoresis, the separation in the gel is very slow, taking several hours and further time is wasted in subsequently extracting (eluting) the separated molecules from the gel. A further problem arises in that molecules tend to elongate and denature when separated in a gel in comparison with a zonal electrophoresis separation. [0004] Earlier electrophoretic separation apparatus and methods have been developed for processing large sample volumes and are not suitable to treat small volumes. Furthermore, the ratio of sample to electrophoresis membrane surface area is usually greater than 2.5 mL/cm.sup.2, typically around 5 mL/cm.sup.2, which results in large dead volumes and the need to re-circulate buffers and samples to reduce heating and prevent clogging of membranes and other separation media. [0005] The present invention seeks to alleviate these problems and in particular, to provide a separation apparatus and methods suitable for use with relatively small sample volumes. SUMMARY OF THE INVENTION [0006] In accordance with the present invention, there is provided a separation apparatus and methods suitable for use with relatively small sample volumes and can effectively and efficiently separate selected products from such small sample volumes. [0007] Further, in accordance with the present invention, there is provide an apparatus for processing compounds in small volumes by electrophoretic separation, the apparatus comprising: [0008] (a) a cathode in a static cathode buffer zone; [0009] (b) an anode in a static anode buffer zone, the anode disposed relative to the cathode so as to be adapted to generate an electric field in an electric field area therebetween upon application of a voltage potential between the cathode and anode; [0010] (c) a first separation barrier disposed in the electric field area; [0011] (d) a second separation barrier disposed between a selected one of the cathode buffer zone and the anode buffer zone and the first barrier so as to define a first interstitial volume therebetween; [0012] wherein in use, electrophoretic buffer is disposed in the cathode buffer zone and the anode buffer zone, a sample constituent is provided to the first interstitial volume; wherein upon application of the voltage potential, a selected separation product is removed from the sample constituent through a selected one of the first and second separation barriers, and provided to a selected one of the cathode buffer and anode buffer zones; and wherein there is substantially no circulation of buffer or sample constituent in the buffer zones or the first interstitial volume. [0013] Still further, in accordance with the present invention, there is provided an apparatus for processing compounds in small volumes by electrophoretic separation, the apparatus comprising: [0014] (a) a cathode in a static cathode buffer zone; [0015] (b) an anode in a static anode buffer zone, the anode disposed relative to the cathode so as to be adapted to generate an electric field in an electric field area therebetween upon application of a voltage potential between the cathode and anode; [0016] (c) a first separation barrier disposed in the electric field area; [0017] (d) a second separation barrier disposed between the cathode buffer zone and the first barrier so as to define a first interstitial volume therebetween; [0018] (e) a third separation barrier disposed between the anode buffer zone and the first barrier so as to define a second interstitial volume therebetween; [0019] wherein in use, electrophoretic buffer is disposed in the cathode buffer zone, the anode buffer zone and at least one of the first interstitial and second interstitial volumes, a sample constituent is provided to a selected one of the first interstitial and second interstitial volumes; wherein upon application of the voltage potential, a selected separation product is removed from the sample constituent, through the first separation barrier, and provided to the other of the first interstitial and second interstitial volumes; and wherein there is substantially no circulation of buffer or sample constituent in the buffer zones, the first interstitial volume or the second interstitial volume. [0020] Preferably, the small volume is less than about 5 mL, preferably about 2 mL or less. The invention is particularly suitable for separating or processing samples of about 0.02 mL to about 0.1 mL. [0021] A ratio of sample to membrane surface area of less that about 1 mL/cm.sup.2 is useful for the present invention. Preferably, the ratio is about 0.5 mL/cm.sup.2 or less, more preferably the ratio is about 0.1 mL/cm.sup.2 or less, and more preferably about 0.02 mL/cm.sup.2. [0022] Through out this specification, the term "buffer" has been used which is intended to include solutions of electrolytes. It will be appreciated that any solution or solvent containing an electrolyte would fall within the definition of "buffer" for the present application. Importantly, the buffer must be a solution which can conduct electricity. Preferably, the solution of electrolytes or buffers used for the present invention have some buffering capacity characteristic of traditional buffers. [0023] An advantage of the present invention is the separation apparatus effectively and efficiently separates selected products from small sample volumes. [0024] Another advantage of the present invention is the ability to load very small samples and carry out fast separations without significant loss of the sample or undue dilution of the samples. [0025] Another advantage of the present invention is the ability to carry out dialysis on very small samples. [0026] These and other advantages and benefits of the invention will be apparent to those skilled in the art upon reading and understanding of the following detailed description. [0027] Throughout this specification, unless the context requires otherwise, the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps. [0028] 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. BRIEF DESCRIPTION OF THE DRAWINGS [0029] FIG. 1 is a schematic drawing illustrating the basic principal of the first aspect of the present invention having two membranes. [0030] FIG. 2 is a schematic drawing illustrating the basic principal of the second aspect of the present invention having three membranes forming a number of compartments and chambers. [0031] FIG. 3 is a schematic drawing illustrating a variant of the second aspect of the present invention. [0032] FIG. 4 is a schematic drawing illustrating an other variant of the basic principal shown in FIG. 3 having a plurality of chambers. Continue reading... Full patent description for Small separation apparatus Brief Patent Description - Full Patent Description - Patent Application Claims Click on the above for other options relating to this Small separation apparatus patent application. ###  How KEYWORD MONITOR works... a FREE service from FreshPatents1. Sign up (takes 30 seconds). 2. Fill in the keywords to be monitored. 3. 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