Compositions and methods for detecting nitration of PKG-1α and methods of use thereof
View Patent ↗Antibodies for detecting nitration of nitrotyrosine 247 PKG-1α and antibodies for detecting nitrotyrosine 425 of PKG-1α are disclosed. Methods of detecting nitrotyrosine 247 PKG-1α and nitrotyrosine 425 of PKG-1α, and uses thereof are also disclosed for identification and diagnosis or phenotypes, pathologies, diseases and disorders associated with protein nitration of PKG-1α are also disclosed. In a preferred embodiment, one or more of the disclosed antibodies is used in the disclosed methods.
1. An isolated monoclonal antibody or antigen binding fragment thereof for detecting nitrated PKG-1α comprising an antigen binding site that binds specifically to an epitope within the protein encoded by the amino acid sequence of SEQ ID NO:4, wherein the epitope comprises the nitrated tyrosine 247 of SEQ ID NO:4.
2. The monoclonal antibody or antigen binding fragment thereof of claim 1 wherein the epitope is a conformational epitope or a linear epitope.
3. The monoclonal antibody or antigen binding fragment thereof of claim 1 , wherein the antibody or antigen binding fragment thereof is a single chain antibody.
4. The monoclonal antibody or antigen binding fragment thereof of claim 1 , wherein the antibody or antigen binding fragment thereof is humanized.
5. The monoclonal antibody or antigen binding fragment thereof of claim 1 , wherein the antigen binding fragment is an Fv, Fab, Fab′, or F(ab′) 2 .
6. The monoclonal antibody or antigen binding fragment thereof of claim 1 , wherein the antibody or antigen binding fragment thereof is bispecific.
7. The monoclonal antibody or antigen binding fragment thereof of claim 1 , wherein the antibody or antigen binding fragment thereof further comprises a radioisotope, a fluorescent compound, a bioluminescent compound, biotin, a chemiluminescent compound, a metal chelator or an enzyme.
8. A method of detecting nitrated PKG-1α comprising detecting nitration of PKG-1α at position Y247 of SEQ ID NO:4 in a biological sample by contacting the biological sample with the monoclonal antibody of claim 1 , wherein the monoclonal antibody of claim 1 comprises a detectable label; and detecting the detectable label.
9. The method of claim 8 wherein the detecting is carried out using mass spectrometry or an immunoassay.
10. The method of claim 8 wherein the immunoassay is selected from the group consisting of radioimmunoassays, ELISAs, immunoprecipitation assays, Western blot, fluorescent immunoassays, and immunohistochemistry.
11. A kit comprising the monoclonal antibody or antigen binding fragment thereof of claim 1 .