The LpMab-23 monoclonal antibody specifically recognizes a cancer-specific form of human podoplanin. Podoplanin is a 38-44 kDa type I transmembrane glycoprotein that is encoded by PDPN. This heavily glycosylated mucin type protein is named for its expression on kidney glomerular epithelial cells known as podocytes. Although the podoplanin protein is expressed on a wide variety of cell types in normal tissues (including placenta, lung, skeletal muscle and brain), LpMab-23 recognizes an altered glycosylation pattern that occurs on oral cancer cells and it shows minimal reactivity with the surrounding non-cancerous tissue. In contrast, the LpMab-17 monoclonal antibody recognizes a non-glycosylated epitope of podoplanin on normal and cancer cells. Podoplanin induces platelet aggregation via its three platelet aggregation-stimulating (PLAG) domains; it binds C-type lectin domain family 1 member B (Clec1B, also known as CLEC2); and it is involved in actin cytoskeleton organization and in cellular adhesion and migration. It also plays roles in organogenesis, vascular development, inflammatory diseases, tumorigenesis, and cancer cell motility and metastasis.
The antibody was conjugated to BD Horizon™ BUV737 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 737-nm. BD Horizon Brilliant BUV737 can be excited by the ultraviolet laser (355 nm) and detected with a 740/35 filter. Due to the excitation of the acceptor dye by other laser lines, there may be significant spillover into channels detecting Alexa Fluor® 700-like dyes (eg, 712/20-nm filter).
Due to spectral differences between labeled cells and beads, using BD™ CompBeads can result in incorrect spillover values when used with BD Horizon BUV737 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different BUV737 reagents (eg, CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone specific compensation controls when using these reagents.