The 5D12 monoclonal antibody specifically recognizes CD366 which is also known as TIM-3 (T-cell immunoglobulin and mucin domain-containing 3) or T-cell membrane protein 3. TIM-3 is encoded by Havcr2 (hepatitis A virus cellular receptor 2). TIM-3 is a type I transmembrane glycoprotein that belongs to the human TIM family within the immunoglobulin superfamily, having one Ig-like V-type domain in its extracellular region. TIM-3 is expressed on activated monocytes, macrophages, dendritic cells, microglia, and mast cells. It is also expressed on type-1 CD4+ (Th1-like) T cells, CD8+ T cytotoxic cells, regulatory T cells (Treg), and natural killer (NK) cells. TIM-3 functions as an inhibitory receptor which helps cells maintain immunological homeostasis and self tolerance. Crosslinking of cell surface TIM-3 by Galectin-9 binding downregulates Th1-like and CD8+ T cell responses and can promote Treg or myeloid-derived suppressor cells. TIM-3 enables dendritic cells to bind phosphatidyl serine expressed by apoptotic cells and to phagocytize these cells to quell potential inflammation. TIM-3 also binds to the alarmin HMGB1 thereby preventing Toll-like receptor (TLR) by released tumor cell DNA.
The antibody was conjugated to an oligonucleotide that contains an antibody clone-specific barcode (ABC) flanked by a poly-A tail on the 3' end and a PCR handle (PCR primer binding site) on the 5' end. The ABC for this antibody was designed to be used with other BD AbSeq oligonucleotides conjugated to other antibodies. All AbSeq ABC sequences were selected in silico to be unique from human and mouse genomes, have low predicted secondary structure, and have high Hamming distance within the BD AbSeq portfolio, to allow for sequencing error correction and unique mapping. The poly-A tail of the oligonucleotide allows the ABC to be captured by the BD Rhapsody™ system. The 5' PCR handle allows for efficient sequencing library generation for Illumina sequencing platforms.
NOTE: The BD Rhapsody Single-Cell Analysis System must be used with the BD Rhapsody Express Instrument.