Application of heterobifunctional ligands in affinity chromatography
Department of Biochemistry, Faculty of Science, Masaryk University, Brno
A very important step in the formation of affinity sorbent is the choice of an appropriate method for immobilization of the affinity ligand. Covalent immobilization usually leads to a statistical orientation of ligands and a certain percentage of the ligands is not accessible for binding. An alternative to covalent immobilization is reversible immobilization that leads to orientation of affinity ligands. Moreover the immobilization can be reversed and the column after regeneration can be employed in an immobilization of different ligand and used in other application. One of possibilities for reversible immobilization is using so called "heterobifunctional ligands". The heterobifunctional ligands are molecules having two binding site - first for the molecule to be purified and second for the solid used as an affinity support. The basic principles of the usage of heterobifunctional ligands in affinity chromatography are given together with the their applications in protein purifications.
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