BcMag™ Glycoprotein & Antibody Conjugation Kit (I)

In their natural condition, most biological compounds do not contain carbonyl ketones or aldehydes. However, similar groups on proteins may be effective for forming an immobilization site that drives covalent coupling away from active centers or binding regions. Glycoconjugates, such as glycoproteins and glycolipids, typically contain sugar residues with hydroxyls on neighboring carbon atoms; these cis-diols can be oxidized with sodium periodate to produce aldehydes as covalent immobilization sites.

BcMag™ Glycoprotein & Antibody Conjugation Kit produces affinity support with glycoproteins and antibodies, which often have numerous carbohydrates (glycosylation) on their Fc sections, using hydrazide-activated magnetic beads and a specific catalyst. These kits may immobilize glycosylated proteins (including monoclonal antibodies) with periodate-oxidizable carbohydrate groups to create reusable affinity matrices. This hydrazide immobilization chemistry results in antibodies with unobstructed antigen-binding sites and excellent purification capacity. In a simple non-amine buffer containing the catalyst, the full coupling reaction takes 2 to 4 hours. Coupling efficiency with antibodies and common glycoproteins is typically greater than 85%, yielding 15μg to 20 μg of immobilized protein per mg of the hydrazide-terminated magnetic beads. The matrix prepared with stable glycoproteins or antibodies can be regenerated and reused at least five times without appreciable loss of binding capacity.
BcMag™ Hydrazide-Terminated Magnetic Beads are uniform magnetic beads grafted with a high density of hydrazide functional groups on the surface. Hydrazide chemistry is effective for labeling, immobilizing, or conjugating glycoproteins via glycosylation sites, which are frequently (as with most polyclonal antibodies) positioned in domains distant from the critical binding sites whose function needs to be preserved. Coupling antibodies in this manner selectively targets heavy chains in the Fc portion of the molecule, assisting in the best possible preservation of antigen binding activity by the ends of the Fv regions. At pH 5 to 7, hydrazide-terminated supports and compounds will conjugate with carbonyls of oxidized carbohydrates (sugars), producing hydrazone linkages. The hydrazide beads are suitable for conjugating larger glycoproteins and glycolipids, carbohydrates or other ligands. Moreover, the hydrophilic surface ensures low nonspecific adsorption, excellent dispersion, and easy handling in various buffers.

Workflow
The Beads work perfectly as affinity resin for affinity purification to refine molecules, cells, and parts of cells into purified fractions. After conjugation with ligands, add the beads to a sample containing the target molecules, then mix, incubate, wash and elute the target molecules

Features and benefits
• Specific immobilization—the hydrazide-activated beads bind exclusively to pure glycoproteins containing sugar groups that have been gently oxidized with periodate (e.g., sialic acid).
• Stable covalent bond with low levels of ligand leakage
• Maintains antibody function – Immobilizes IgG via the Fc region, leaving both antigen binding sites available for target capture.
• Low nonspecific binding
• High capacity – Immobilize 15-20μg antibody/mg beads

Please contact us at for specific academic pricing.