Construction of phosphorylatable monoclonal antibody CC49 with a casein kinase II recognition site.
A phosphorylation site for casein kinase II was introduced into chimeric monoclonal antibody CC49 (MAb-chCC49) by site-specific mutation of the coding sequence. The phosphorylation site for the casein kinase II was positioned at the carboxyl terminus of the heavy chain constant region of the MAb-chCC49. The resultant modified MAb-chCC49CKII was expressed in NS0 cells and purified. The MAb-chCC49CKII protein was phosphorylated by casein kinase II with [gamma-32P]ATP to high radiospecific activity. The 32P-labeled MAb-chCC49CKII binds to cells expressing TAG-72 antigens. The introduction of the phosphorylation sites for casein kinase II into monoclonal antibodies (MAb) provides a new reagent for the diagnosis and treatment of cancers. This demonstrates that the casein kinase II recognition site can also be used to introduce phosphorylation sites into proteins.
Lin
L
L
Department of Molecular Genetics and Microbiology, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway 08854-5635, USA.
Gillies
S D
SD
Schlom
J
J
Pestka
S
S
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000378:metabolism; D000912:Antibodies, Neoplasm / Q000097:blood / Q000737:chemistry* / Q000378:metabolism*; D001483:Base Sequence; D047390:Casein Kinase II; D002417:Cattle; D017931:DNA Primers; D006801:Humans; D007127:Immunoglobulin Constant Regions / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D016297:Mutagenesis, Site-Directed; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D015183:Restriction Mapping; D013379:Substrate Specificity; D014162:Transfection; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000912:Antibodies, Neoplasm; C091202:B72.3 antibody; D017931:DNA Primers; D007127:Immunoglobulin Constant Regions; D011994:Recombinant Proteins; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases