As a leading global company, Creative Biolabs has established an advanced platform to offer the most comprehensive glycan modification and labeling service. We own a team of experts who have accumulated extensive experience in the biotinylation of diverse glycans. We are pleased to tailor the most appropriate strategies for biotinylation to meet every demand of our customers.
Biotin is widely utilized in biotechnological applications as a labeling reagent or affinity purification tag owing to its extremely high affinity. It was first exploited in immunocytochemical applications in the mid-1970s and has since been commonly used to localize antigens in cells and tissues. Generally, biotinylation is rapid, specific, and unlikely to perturb the natural function of the molecule due to the small size of biotin. The labeled molecule then can be detected in enzyme-linked immunosorbent assay (ELISA), dot blot or western blot (WB) methods using streptavidin or avidin probes. Therefore, it is widely used in the study of glycoproteins and glycan-binding proteins.
Since biotinylation is rapid, specific, and is unlikely to perturb the natural function, it is popularly used in the study of interactions. Creative Biolabs has accumulated years of experience in carbohydrate biotinylation service, we are confident in offering high-quality custom biotinylation services according to our customers' specific demands. Different strategies can be employed for biotinylation including but not limited to N-hydroxysuccinimide (NHS), enzymatic biotinylation with E. coli biotin ligase (BirA), and proximity-dependent biotin identification (BioID).
NHS ester-activated biotins are the most popular type of biotinylation reagent. NHS esters react efficiently with primary amino groups (-NH2) in pH 7-9 buffers to form stable amide bonds. Because glycoprotein and other proteins generally contain multiple lysine (K) residues in addition to the N-terminus of each polypeptide, they have multiple primary amines available as targets for labeling with NHS-activated reagents.
BirA binds biotin and ATP and generates the intermediate, biotinoyl-5 -AMP (bioAMP). At low biotin concentrations, bioAMP is transferred to the -NH2 of a lysine in BCCP. Under conditions of high biotin concentration, where all BCCP subunits are biotinylated, bioAMP remains bound to BirA; in vitro the BirA-bioAMP complex has a half-life of ~30 min. In bacteria, the BirA-bioAMP complex binds to a 40-base pair sequence in the biotin biosynthetic operon, resulting in the transcriptional repression of genes encoding enzymes required for biotin biosynthesis.
BioID is a unique method to screen for physiologically relevant protein interactions that occur in living cells. Based on its mechanism of action, BioID is effective when applied to insoluble and membrane-associated proteins, two classes of proteins that may be refractory to screening with conventional approaches. Additional advantages include the ability to identify weak and/or transient interactions, the ability to screen for interactions in a relatively natural cellular setting, and temporal inducibility of biotin labeling. With modifications, BioID is effective in a wide variety of cell types and species.
Fig.1 Coupling of biotin-LC-hydrazide to the reducing ends of carbohydrates. (Grün, 2006)
Creative Biolabs is a forward-looking company that has devoted to biotinylation services for many years. Strong scientists team and experienced technical support make us confident in providing the best glycan modification and labeling services to global customers. Please feel free to contact us for more information.