Method and apparatus determining the isoelectric point of charged analyte

• US 20070163884A1
• Filed: 01/12/2004
• Published: 07/19/2007
• Est. Priority Date: 01/15/2003
• Status: Active Grant

• Current Assignee: Protasis Corp.
• Original Assignee: Protasis Corp.
• Inventors: Strand, David, Leatzow, Dan
• Granted Patent: US 8,080,145 B2
• Time in Patent Office: Days
• US Class Current: 204/548
• CPC Class Codes: C07K 1/28 Isoelectric focusing
• CPC Class Codes: G01N 27/44704 Details; Accessories

Abstract

Devices are provided for determining the isoelectric point of a charged analyte, comprising a titration chamber and an electrode chamber. The electrode chamber comprises at least two electrodes, for example, an electrode array. Either or both of the titration chamber and the electrode chamber may have a shaped geometry. The electrodes are operative, in conjunction with the shaped geometry of the chamber(s) where appropriate, to generate an electric field gradient in the titration chamber. Permeable material separates the titration chamber and the electrode chamber. A pH Sensor is located in the titration chamber for obtaining the pH of the first fluid. Certain preferred embodiments further include an analyte band detector for detecting the presence and optionally the location of a focused band of charged solute. Methods are provided for determining the isoelectric point of a charged analyte comprising introducing a carrier fluid comprising a Charge analyte into the titration chamber of a device as just described and applying an electric field gradient to focus the charged analyte into a focused band. The pH of the carrier fluid is incremented or adjusted to shift the location of the focused band of charged analyte, and the pH and location of the focused band of charged analyte are obtained for a plurality of locations and pH'"'"'s and the isoelectric point is determined from such data.

Claims

1-19. -19. (canceled)

20. A method of determining the isoelectric point of a charged analyte comprising:

focusing a charged analyte in a flowing liquid in an electric field gradient to form in the flowing liquid a focused band of the charged analyte at a stable position in the electric field gradient;

changing the pH of the flowing liquid at least once by an amount sufficient to change the position of the focused band of the charged analyte within the electric field gradient;

obtaining pH and corresponding position data for the charged analyte, comprising determining the pH of the flowing liquid and the corresponding position of the focused band of the charged analyte at a plurality of band positions within the electric field gradient; and

determining the isoelectric point of the charged analyte based on the pH and corresponding position data.

21-97. -97. (canceled)