An anti-human ppGalNAcT-2 monoclonal antibody

Aberrant mucin O-glycosylation is a pathological alteration that is widespread in cancer. The UDP-N-acetyl-D-galactosamine polypeptide N-acetylgalactosaminyltransferases (ppGalNAc-T) family of enzymes regulates the initial key steps of mucin O-glycosylation. ppGalNAc-T2, as a member of the ppGalNAc-T family, was recently described as an altered expression in oral squamous cell carcinoma and colorectal and breast carcinoma. In order to gain further insight into the role of ppGalNAc-T2, we produced the anti-human ppGalNAc-T2 monoclonal antibody (MAb) 5F3. The IgM κ isotype of this MAb was further characterized using ELISA, Western blot analysis, flow cytometry, and immunofluorescent staining. In the present study, MAb 5F3 can specifically recognize human ppGalNAc-T2 protein in various formats by Western blot analysis, flow cytometry, and immunofluorescent staining. For the first time, it was shown that both Jurkat and HepG2 cell lines clearly express ppGalNAc-T2.