Electrophoretically mediated chemical analysis

• US 5,810,985 A
• Filed: 02/09/1995
• Issued: 09/22/1998
• Est. Priority Date: 09/14/1992
• Status: Expired due to Fees

• Current Assignee: Purdue Research Foundation (Purdue University)
• Original Assignee: Purdue Research Foundation (Purdue University)
• Inventors: Bao, Jianmin, Harmon, Bryan Joseph, Patterson, Dale Hansen, Regnier, Fred E.
• Primary Examiner(s): Tsang, Cecilia J.
• Assistant Examiner(s): DELACROIX MUIRHEI, CYBILLE
• Application Number:
  US08/386,224
• Time in Patent Office: 1,321 Days
• Field of Search: 204/182.1, 204/182.8, 204/299 R, 204/451, 204/452, 204/601, 204/603, 435/7.4
• US Class Current: 204/451
• CPC Class Codes: G01N 27/44743 Introducing samples
• CPC Class Codes: G01N 27/44747 Composition of gel or of carrier mixture

Abstract

Disclosed are methods of electrochemical analysis of an analyte in a sample. The methods include providing a sample comprising an analyte, and introducing the sample into a capillary tube containing electrophoretic running buffer and a reactant. According to the methods of the invention, one or both of the analyte and the reactant is electrically charged. Chemical contact between analyte and reactant induced by their relative electrophoretic mobility results in the breaking or formation of a covalent bond of one of the analyte or the reactant to produce a detectable product. The methods also include the steps of imposing along the length of the capillary tube an electric current for a time sufficient to bring into chemical contact within the tube analyte and reactant so as to allow formation of product, and detecting the product.

Claims

1. A method of analysis of an analyte in a sample, the method comprising:

(a) introducing first and second reactants and a sample comprising an analyte into a capillary containing an electrophoretic running buffer, wherein chemical contact between said analyte and said first reactant results in the breaking or formation of a covalent bond in said analyte to produce a first product, and wherein chemical contact between said first product and said second reactant results in the breaking or formation of a covalent bond in said first product to produce a detectable second product, and wherein said analyte and said first product or said reactants are electrically charged;

(b) imposing along the length of said capillary an electric potential for a time sufficient to bring into chemical contact by electrophoretic mobility within said capillary said analyte and said first reactant so as to induce formation of said first product, and then said first product and said second reactant to produce said second detectable product; and

(c) detecting said second product.

2. The method of claim 1 wherein said capillary used in step (a) contains said first and second reactants loaded thereinto prior to the introduction of said sample.

3. The method of claim 1 wherein said electric potential is imposed on said capillary for a time sufficient to separate said second product from said analyte or said reactants prior to step (c).

4. The method of claim 1 wherein said electric potential is imposed on said capillary for a time sufficient to separate by electrophoretic motion said analyte from other sample components prior to said chemical contact between said analyte and said first reactant.

5. The method of claim 1 wherein said analyte and said first product comprise an enzyme substrate and said first and second reactants each comprise an enzyme.