Matrixes, arrays, systems and methods
View Patent ↗The present invention relates to matrixes, arrays, systems and methods for analyzing biomolecules by their isoelectric point, optionally, in combination with a second dimension analysis. The assortment of matrixes, arrays and systems provided herein are useful for causing a biomolecule under the influence of an electrical field to accumulate into an buffer that comprises a pH value that is the same as the isoelectric point of the biomolecule. The methods of this invention are useful for, e.g., research and diagnostic purposes.
1. A method for sorting a mixture of biomolecules comprising the steps of adding the mixture to a running buffer in a system for isoelectric focusing (IEF) in a matrix comprising at least two compartments or cells that are not adjoined, the compartments or cells containing a first IEF buffer and at least a second different IEF buffer, and a running buffer that circulates through and around the matrix delivering the mixture to the compartments, and applying a current to the running buffer until the biomolecules in the mixture are separated into the compartments.
2. The method of claim 1 for diagnosing or prediagnosing a disease state in a subject comprising the steps of:
(1) adding a sample from the subject to the running buffer in the system, generating an electrical field, and circulating the running buffer across the IEF buffer; and
(2) comparing the biomolecules in the compartments containing IEF buffers from the subject to the biomolecules from a sample from a normal subject that does not have the disease or is not predisposed to having the disease.
3. The method according to claim 1 , wherein the biomolecules from the normal subject are prepared by adding a sample from the normal subject to the running buffer, generating the electrical field and circulating the running buffer across the IEF buffer.
4. The method according to claim 1 , further comprising the step of subjecting the biomolecule to a second dimension analysis.
5. The method according to claim 4 , wherein the second dimension analysis separates the biomolecules by the mass of the biomolecule.
6. The method according to claim 5 , wherein the second dimension analysis is by polyacrylamide gel electrophoresis (PAGE).
7. The method according to claim 5 , wherein the second dimension is by Mass spectrometry, capillary electrophoresis, and liquid chromatography.
8. The method according to claim 4 , further comprising the step selected from the group consisting of:
(a) comparing the location of the biomolecule in the second dimension analysis to another biomolecule in the second dimension analysis;
(b) determining the amount of the biomolecule in the second dimension analysis.
9. The method of claim 1 for sorting a biomolecule in a sample comprisinexposing the sample to an alternating electrical field.