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.

Description

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 membrane...

AI Technical Summary

Benefits of technology

Enables fast and efficient separation of small samples with minimal loss and dilution, suitable for volumes as low as 0.02 mL to 0.1 mL, and facilitates dialysis without recirculation, reducing heating and maintaining sample integrity.

Problems solved by technology

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.

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.

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 / cm2, typically around 5 mL / cm2, 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.

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