Immunoaffinity isolation of modified peptides from complex mixtures

• Main Authors: ZHA, XIANGMING, ZHANG, HUI, COMB, MICHAEL J, TAN, YI, RUSH, JOHN
• Format: Patent
• Language: English; French; German
• Published: 11.01.2012
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PHYSICS; PROCESSES OF PREPARING SUCH COMPOSITIONS; SPIRITS; TESTING; VINEGAR; WINE

The invention is a method for isolating a target population of naturally-occurring post-translationally modified peptides from a complex mixture of peptides, said method comprising the steps of: (a) obtaining a digested proteinaceous preparation from an organism, wherein said digested proteinaceous preparation comprises a complex mixture of peptides comprising naturally-occurring post-translationally modified peptides from two or more different proteins and further fractionating said post-translationally modified peptides in said digested proteinaceous preparation by reverse-phased chromatography to produce a fractionated proteinaceous preparation; (b) contacting said fractionated proteinaceous preparation with at least one immobilized post-translational modification-specific antibody; and (c) isolating said target of naturally-occurring post-translationally modified peptides specifically bound by said immobilized modification-specific antibody in step (b).