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™ BUV563 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 which has an Ex Max of 348 nm and an acceptor dye. The tandem has an Em Max at 563 nm. BD Horizon BUV563 can be excited by the 355 nm ultraviolet laser. On instruments with a 561 nm Yellow-Green laser, the recommended bandpass filter is 585/15 nm with a 535 nm long pass to minimize laser light leakage. When BD Horizon BUV563 is used with an instrument that does not have a 561 nm laser, a 560/40 nm filter with a 535 nm long pass may be more optimal. Due to the excitation and emission characteristics of the acceptor dye, there may be spillover into the PE and PE-CF594 detectors. However, the spillover can be corrected through compensation as with any other dye combination.