Rapid quantitative analysis of proteins or protein function in complex mixtures

US6670194B1 · kind B1 · utility

Assignee

• University of Washington · US

Inventors

• Rudolf Aebersold · Zürich, CH
• Michael H. Gelb · Seattle, US
• Steven P. Gygi · Cambridge, US
• C. Ronald Scott · Seattle, US
• Frantisek Turecek · Seattle, US
• Scott Gerber · Seattle, US
• Beate Rist · Giesen, DE

Key dates

• Filing date: Aug 25, 1999
• Grant date: Dec 30, 2003
• Expiry date: Aug 25, 2019

Classification

• Technology area (CPC Y): Emerging Cross-Sectional Technologies
• CPC primary: Y10T436/25375
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

Analytical reagents and mass spectrometry-based methods using these reagents for the rapid, and quantitative analysis of proteins or protein function in mixtures of proteins. The methods employ affinity labeled protein reactive reagents having three portions: an affinity label (A) covalently linked to a protein reactive group (PRG) through a linker group (L). The linker may be differentially isotopically labeled, e.g., by substitution of one or more atoms in the linker with a stable isotope thereof. These reagents allow for the selective isolation of peptide fragments or the products of reaction with a given protein (e.g., products of enzymatic reaction) from complex mixtures. The isolated peptide fragments or reaction products are characteristic of the presence of a protein or the presence of a protein function in those mixtures. Isolated peptides or reaction products are characterized by mass spectrometric (MS) techniques. The reagents also provide for differential isotopic labeling of the isolated peptides or reaction products which facilitates quantitative determination by mass spectrometry of the relative amounts of proteins in different samples. The methods of this invention …