Patent Number: 6,309,863

Title: Methods for generating phosphorylation site-specific immunological reagents

Abstract: The present invention provides methods for generating phosphorylation site-specific immunological reagents. More specifically, a phosphopeptide mimetic is incorporated into a polypeptide in place of a phosphorylated amino acid. The polypeptide is used as antigen by standard methods to generate either monoclonal or polyclonal antibodies which cross-react with the naturally phosphorylated polypeptide. The phosphopeptide mimetic preferably contains a non-hydrolyzable linkage from the appropriate carbon atom of the amino acid residue to a phosphate group. A preferred linkage is a CF.sub.2 group. Such a linkage is used to generate the phosphoserine mimetic F.sub.2 Pab, which is incorporated into a polypeptide sequence derived from p53 to produce antibodies which recognize a specific phosphorylation state of p53. A CF.sub.2 group linkage is also used to produce the phosphothreonine mimetic F.sub.2 Pmb, and to produce the phosphotyrosine mimetic, F.sub.2 Pmp.

Inventors: Anderson; Carl W. (Stony Brook, NY), Appella; Ettore (Montgomery, MD), Sakaguchi; Kazuyasu (Montgomery, MD)

Assignee: Brookhaven Science Associates

International Classification: C07K 16/32 (20060101); C07K 16/18 (20060101); C07K 016/00 (); C12P 021/04 ()

Expiration Date: 10/30/2018