Pat. No.: 5,006,210; Date: Apr. 9, 1991; Lab: Ames Lab.
Inv.: Yeung, E.S; Kuhr, W.G.
The Microfluor microfluorophotometric capillary zone electrophoresis detector measures laser-excited fluorescence of chemical and biological molecules at unprecedented high speeds and low levels, including single molecules and cells, and is useful in biological and chemical detection and analytical instruments. Apparatus and methods are provided for capillary zone electrophoresis with laser-induced indirect fluorescence detection. The detector can analyze with higher sensitivity samples 50-times smaller than those required by other fluorescence detectors at concentrations as low as 10(-11) molar (10 parts per trillion), including detection and quantification of samples subjected to capillary electrophoresis. The method and apparatus can increase the rate of gene sequencing by several orders of magnitude--genomes that would require 100s of years to sequence with current technology can be sequenced in a matter of weeks with 99% accuracy. The detector can also be used for protein sequencing and detection of most biochemical and other reactions involving fluorescent reactants. Using fluorescent tagged nucleotides or amino acids, the Microfluor detector can be used for gene and protein sequencing. The detectorÕs ability to study chemical activity within a single cell can be used to screen and study the effects of drugs, toxic agents and carcinogens.This is the first laser-based microfluorophotometric detector. Unlike other laser-excited detectors, this is not affected by stray light and can be operated in a fully lighted room. A detector is positioned on the capillary tube of a capillary zone electrophoresis system. The detector includes a laser which generates a laser beam which is imposed upon a small portion of the capillary tube. Fluorescence of the elutant electromigrating through the capillary tube is indirectly detected and recorded.
Status - Exclusively licensed in July 1995 to Premier American Technologies Corp. (PATCO; Bellefonte, PA) for gene sequencing and other biotechnology uses. Previously, exclusively licensed to Lachat Instruments, Inc. (Milwaukee, WI) for water pollution monitoring-related fields of use. Patent assigned to Iowa State University Research Foundation, Inc. as granted (contractor operator of Ames Labs.), but has been repeatedly reported as a DOE invention.
PATCO, a manufacturer and worldwide marketer of analytical instrumentation, will use this technology to develop detectors that will increase the speed of gene sequencing several orders of magnitude. While it takes current gene sequencers about 1,000 or more years to sequence the approximately 3 billion base pairs in the human genome, new sequencers using this technology will ba able to do this in about 68 days. PATCO expects to introduce Microflour-based gene sequencing apparatus in 1996.Researchers at Ames Lab. have been using the detector to study adduct formation and base modification in DNA and the effects of carcinogens within cells, the chemical content of single cells to test for drug and toxic effects, and for development of cancer diagnostics. This invention was discussed in, "Indirect Detection Methods for Capillary Separations," Analytical Chemistry, vol. 63, p. 275A-80A, 1991. This technology received a 1991 ÒR&D-100 AwardÓ from Reseach and Development Magazine.