%0 Journal Article
%T Development and characterization of anti©\glycopeptide monoclonal antibodies against human podoplanin, using glycan©\deficient cell lines generated by CRISPR/Cas9 and TALEN
%A Hiroyoshi Suzuki
%A Hiroyuki Harada
%A Michiaki Takagi
%A Mika K. Kaneko
%A Ryusuke Honma
%A Satoshi Ogasawara
%A Shinji Abe
%A Takuro Nakamura
%A Yasuhiko Nishioka
%A Yuki Fujii
%A Yukinari Kato
%J Archive of "Cancer Medicine".
%D 2017
%R 10.1002/cam4.954
%X Human podoplanin (hPDPN), which binds to C©\type lectin©\like receptor©\2 (CLEC©\2), is involved in platelet aggregation and cancer metastasis. The expression of hPDPN in cancer cells or cancer©\associated fibroblasts indicates poor prognosis. Human lymphatic endothelial cells, lung©\type I alveolar cells, and renal glomerular epithelial cells express hPDPN. Although numerous monoclonal antibodies (mAbs) against hPDPN are available, they recognize peptide epitopes of hPDPN. Here, we generated a novel anti©\hPDPN mAb, LpMab©\21. To characterize the hPDPN epitope recognized by the LpMab©\21, we established glycan©\deficient CHO©\S and HEK©\293T cell lines, using the CRISPR/Cas9 or TALEN. Flow cytometric analysis revealed that the minimum hPDPN epitope, in which sialic acid is linked to Thr76, recognized by LpMab©\21 is Thr76¨CArg79. LpMab©\21 detected hPDPN expression in glioblastoma, oral squamous carcinoma, and seminoma cells as well as in normal lymphatic endothelial cells. However, LpMab©\21 did not react with renal glomerular epithelial cells or lung type I alveolar cells, indicating that sialylation of hPDPN Thr76 is cell©\type©\specific. LpMab©\21 combined with other anti©\hPDPN antibodies that recognize different epitopes may therefore be useful for determining the physiological function of sialylated hPDPN
%K Epitope
%K glycopeptide
%K monoclonal antibody
%K podoplanin
%K sialic acid
%U https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5313638/