The Preparation Of Anti-human MUC1 Monoclonal Antibodies And Expression Of Glycosylated MUC1

MUC1 is a polymorphic, highly glycosylated type I transmembrane protein, normally found at the apical surface of mucin-secreting epithelial cells in many organs including pancreas, breast, airway and gastrointestinal tract. MUC1 contains carbohydrate content and core polypeptide, the bulk of glycosylation, which can make up between 50 and 90% of the total mass, is O-linked to numerous threonine and serine residues in the core polypeptide;the core polypeptide has three parts including an extracellular region, a small transmembrane region and an intracellular tail. The extracellular region essentially consists of 20-120 viariable number tandem repeats (VNTRs) wherein each copy of VNTRs contains 20 amino acids with a sequence of VTSAPDTRPAPGSTAPPAHG. In addition, each VNTR comprises five potential O-glycosylation sites. It has been found that most of the antibodies elicited against MUC1 targets to VNTR, mainly to its hydrophilic region (PDTRPAP). In normal epithelial cells, the VNTR is densely coated with oligosaccharides. However, MUC1 derived-tumor cells is much less glycosylated than normal MUC1 protein, resulting in the exposure of normally cryptic peptide core and new carbohydrate epitopes. Accumulating evidence shows that aberrantly glycosylated MUC1 in malignant cells can shed off into circulation as a tumor marker that aids diagnosis, determines prognosis, prospectively predicts response or resistance to specific therapies.To detect the soluble MUC1, various MUC1 McAbs have been generated to estabilsh ELISA Kits. And yet it is difficult to select the proper immunogen to produce MUC1 McAbs due to the complexity of glycosylation of MUC1. At present, human milk-fat globule membranes (HMFGM), MUC1-expressing tumors, MUC1 mucin-related synthetic peptides and glycopeptides have been used as immunogen. In the research, we try to...