Anti-phospho-Akt antibodies
View Patent ↗The Present invention relates to antibodies that immunospecifically bind to phospho-Akt and certain p-Akt substrates. The invention encompasses human and humanized forms of the antibodies and their use in treating cancers and other proliferative disorders. The invention also relates to p-Akt derived peptides useful for preparing the antibodies. Methods and compositions for detecting, diagnosing, treating or ameliorating a disease or disorder, especially cancer and other proliferative disorders using the present antibodies also are disclosed.
1. A method for detecting the presence or the amount of a phospho-AKT protein in a biological sample comprising,
(a) contacting the sample with a monoclonal antibody which specifically binds to a phosphorylated epitope within the peptide of SEQ ID NO:5 or SEQ ID NO:6; and
(b) determining the presence or the amount of phospho-AKT protein bound to the monoclonal antibody,
wherein the monoclonal antibody is obtained by a method comprising the steps of:
(i) administering to a mammal with at least two peptides, wherein the two peptides consist of the amino acid sequences of SEQ ID NOs: 5 and 6, respectively, and wherein the two peptides are phosphorylated;
(ii) collecting cells producing the antibody from the mammal; and
(iii) immortalizing the cells obtained in step (ii) thereby creating a hybridoma expressing the monoclonal antibody.
2. The method of claim 1 , wherein the monoclonal antibody is labeled with a detectable label.
3. The method of claim 2 , wherein the detectable label is a radiolabel, a fluorescent label, a luminescent label, a bioluminescent label or an enzymatic label.
4. The method of claim 3 , wherein the amount of phospho-AKT protein is determined by an immunoassay.
5. The method of claim 4 , wherein the immunoassay is selected from the group consisting of radioimmunoassay (RIA), enzyme-linked immunoabsorbent assay (ELISA), immunoprecipitation (IP), immunohistochemistry (IHC), and immunofluorescence (IF).
6. The method of claim 5 , wherein the biological sample is obtained from cancerous tissues.
7. The method of claim 1 , wherein the two peptides are phosphorylated in at least one serine or threonine residue within the peptides.