Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Reexamination Certificate
2011-06-28
2011-06-28
Cheu, Jacob (Department: 1641)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
C435S007200
Reexamination Certificate
active
07968299
ABSTRACT:
The present invention is method of determining whether or not associations between a given protein and other proteins in a cell are specific. The method comprises (a) providing a first sample of the cells in which the given protein contains a tag, (b) providing a second sample of the same cells, wherein the given protein and the other proteins are metabolically labeled, and wherein neither the given protein nor the other proteins are tagged, (c) mixing and lysing the first cell sample and the second cell sample to provide a mixture of proteins, (d) binding the tag of the given protein to a substrate, (e) isolating proteins associated with the tagged given protein bound to the substrate, whereby the associated proteins comprise: (i) proteins specifically associated with the tagged given protein, (ii) proteins non-specifically associated with the tagged given protein, or (iii) a combination thereof, (f) determining whether each associated protein is unlabeled or a mixture of labeled and unlabeled proteins, wherein if the associated protein is not labeled, then that protein was specifically associated in the cell with the tagged given protein.
REFERENCES:
Schulze et al. “A Novel Proteomic Screen for Peptide-Protein Interactions.”The Journal of Biological Chemistry, Mar. 12, 2004, 279(11):10756-10764.
Aebersold et al. “Mass Spectrometry-based Proteomics.”Nature, Mar. 13, 2003, 422:198-207.
Ong et al. “Properties of13C-Substituted Arginine in Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC),”Journal of Proteome Research, (2003), 2:173-181 (Published on web Dec. 13, 2002).
Terpe, K., “Overview of Tag Protein Fusions: From Molecular and Biochemical Fundamentals to Commercial Systems.”Applied Microbiology and Biotechnology, Jan. 2003, 60:523-533 (Published online Nov. 7, 2002).
Chait Brian T.
DeGrasse Jeffrey A.
Oeffinger Marlene
Rout Michael P.
Sekedat Matthew D.
Cheu Jacob
Hoffman & Bacon, LLP
The Rockefeller University
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