BcMag™ Glycoprotein & Antibody Conjugation Kit (II)

BcMag™ Glycoprotein & Antibody conjugation kit enables the formation of immobilization sites in biological compounds using carbonyl ketones or aldehydes present in similar groups on proteins. These sites drive covalent coupling away from active centers or binding regions. Glycoconjugates like glycoproteins and glycolipids contain sugar residues with hydroxyls on neighboring carbon atoms. Oxidizing these cis diols with sodium periodate results in aldehydes that serve as covalent immobilization sites. The kit produces affinity support with glycoproteins and antibodies using hydrazide-activated magnetic beads and a specific catalyst. Monoclonal antibodies and glycosylated proteins can be immobilized using periodate-oxidizable carbohydrate groups to create reusable affinity matrices. This hydrazide immobilization chemistry yields antibodies with unobstructed antigen-binding sites and excellent purification capacity. The full coupling reaction takes 2 to 4 hours in a simple non-amine buffer containing the catalyst. Antibodies and common glycoproteins show coupling efficiency 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. The BcMag™ Cleavable Hydrazide-Terminated Magnetic Beads are magnetic beads grafted with a high density of cleavable hydrazide functional groups on the surface. Hydrazide chemistry is effective for labeling, immobilizing, or conjugating glycoproteins via glycosylation sites that are frequently 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. Hydrazide-terminated supports and compounds conjugate with carbonyls of oxidized carbohydrates (sugars) to produce hydrazone linkages at pH 5 to 7. The hydrazide beads are suitable for conjugating larger glycoproteins and glycolipids, carbohydrates, or other ligands. 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.

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