Electrophoresis apparatus and method

Abstract

An electrophoretic apparatus comprising: a first electrolyte chamber containing a first electrode; a second electrolyte chamber containing a second electrode; a first sample chamber disposed between the first and second electrolyte chambers and proximate to the first electrolyte chamber; a second sample chamber disposed between the first sample chamber and the second electrolyte; three ion-permeable barriers separating the first electrolyte chamber, the first sample chamber, the second sample chamber, and the second electrolyte chamber, respectively, wherein the ion-permeable barriers impede convective mixing of the contents in each of the respective chambers; a first electrolyte reservoir and a second electrolyte reservoir in fluid communication with the first and second electrolyte chambers, respectively; a first sample reservoir and a second sample reservoir in fluid communication with the first and second sample chambers, respectively; means adapted for communicating a first electrolyte and a second electrolyte between the respective electrolyte chambers and reservoirs; means adapted for communicating a first fluid and a second fluid between the respective sample chambers and reservoirs, wherein at least one of the first and second fluid contains at least a sample, wherein application of an electric potential causes migration of at least one component through at least one of the ion-permeable barriers.

Description

BACKGROUND OF THE INVENTION [0001] The present invention relates to an electrophoresis apparatus and method suitable for electrophoretically altering the original composition of a mixture that contains at least one ampholytic component. [0002] When ampholytic compounds, such as amino acids, peptides, oligopeptides, proteins, and the like are present in a solution at a low concentration, their charge-state depends on the pH of their environment. At a certain characteristic pH value, the net charge—and consequently, the electrophoretic mobility—of an ampholytic compound becomes zero. That pH value is called the pI value of the ampholytic compound. When two ampholytic compounds have different pH values, their net charge becomes zero at different pH values. Thus, if a pH gradient is established in an electric field, the two ampholytic species achieve zero net charge at different points of the pH gradient that can result in their separation. Such separations are called isoelectric focusi...

AI Technical Summary

Benefits of technology

The present invention provides an electrophoresis apparatus and method that can alter the composition of a mixture containing ampholytic components. The apparatus minimizes the distance the components have to migrate electrophoretically, achieves the electrophoretic separation without causing detrimental heating effects, and maximizes the production rate in unit separation space and time. The apparatus also requires minimal amounts of auxiliary agents and reduces the use of fluid in the system. The apparatus includes an electrolyte chamber and a sample chamber separated by an ion-permeable barrier, which prevents convective mixing of the components. The apparatus can also circulate electrolyte and sample streams separately, allowing for individual control of the movement of each. The separation unit can be a cartridge or cassette fluidly connected to the electrolyte and sample reservoirs.

Problems solved by technology

IEF separations often take considerable time because the electrophoretic mobility of each ampholytic species becomes low as they approach the point in the pH gradient where they become isoelectric.

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