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The preactivated Glyoxal Agarose base beads offered by Agarose Bead Technologies possess all the required properties of preferred supports for the preparation of affinity resins
Affinity chromatography is based on selective binding interaction between an immobilized ligand and the target molecule. Affinity chromatography plays a dominant role in the development of simple purification protocols that can be applied at both research and manufacturing scales.
Affinity chromatography resins are made by immobilization of ligands on the surface of base beads. The type of ligand used will determine the resin selectivity, while the type of beads will determine binding capacities as well as the level of nonspecific adsorption. Agarose Bead Technologies offers a family of preactivated agarose base matrices specifically developed for the preparation of affinity chromatography resins.
These base matrices are preactivated with aldehydes groups and are marketed under the trade name Glyoxal Agarose (GA). Glyoxal chemistry enables the coupling of proteinaceous ligands via reductive amination of the primary amino groups on the ligand surface. This chemistry enables stable conjugation of ligands via multipoint covalent attachment and has been proven advantageous over other immobilization chemistries due to the increased stabilization of the coupled ligand.
"THIS CHEMISTRY ENABLES STABLE CONJUGATION OF LIGANDS VIA MULTIPOINT COVALENT ATTACHMENT"
The high activation level is one of the main advantages of GA support as compared to other types of activated beads. In reductive amination, ligand immobilization is driven by the higher density of reactive groups (primary amino groups, lysins) on the protein surface.
Consequently, ligands are “self-directed” onto the agarose surface by the area where multipoint covalent immobilization is more favorable.
For this reason, GA has been successfully used for the stabilization of enzymes and proteins via multipoint covalent attachment with a low impact on the properties of immobilized ligand. For instance, between 1 to 4-logs improvement in the stability of the immobilized enzymes with at least 70% of their activity have been reported for enzymes immobilized on glyoxal agarose (1).
The preactivated Glyoxal Agarose base beads possess all the required properties of preferred supports for the preparation of affinity resins. The superiority of these beads is associated with the character of glyoxal conjugation chemistry and with the vast selection of particle and pore sizes enabling either high capacities of the target and/or process scenarios.
1. Hussain, Fouzia et al., Molecules 23.12 (2018); Guisan, Jose M. et al., Methods in Molecular Biology. Vol. 2100, (2020), p. 83–92.