METHODS FOR QUANTITATIVE DETERMINATION OF PROTEIN-NUCLEIC ACID INTERACTIONS IN COMPLEX MIXTURES

摘要

In various embodiments, the present invention relates generally to analysis of complex mixtures and, more specifically, to detection and quantitative determination of multiple proteins, protein modifications, and protein-nucleic acid interactions in those complex mixtures.

主权项

1. A method of detecting and quantifying a plurality of unique proteins present in a mixture, at least some of the unique proteins binding in the mixture to nucleic acid fragments, the method comprising the steps of:
providing a plurality of identification vehicles each consisting essentially of (a) a binding portion specific to a unique protein in the mixture, and (b) an oligonucleotide portion; incubating the identification vehicles with the mixture, whereby the binding portions of at least some of the vehicles bind to corresponding proteins in the mixture; ligating the oligonucleotide portions of bound identification vehicles with the nucleic acid fragments bound to proteins to which the binding portions of the identification vehicles are themselves bound, thereby forming complexes each having a protein, a binding vehicle, and a combined nucleic acid strand including the oligonucleotide of the identification vehicle and a protein-bound nucleic acid fragment; and sequencing at least a portion of the combined nucleic acid strands to quantify the proteins corresponding thereto and identify the nucleic acid fragments to which the proteins bind.