Affinity Chromatographic Purification of Antibodies

Abstract

Affinity chromatographic methods for purifying antibodies are reviewed. Topics reviewed include (a) the matrices used in the preparation of the affinity supports; (b) the chemistries commonly used to attach affinity ligands directly on hydroxyl-carrying supports and indirectly through the use of bifunctional agents to crosslink the affinity ligands to the supports; (c) methods for detecting ligand leakage; (d) macromolecular affinity ligands; (e) low molecular weight peptidyl affinity ligands; (f) low molecular weight non-peptidyl affinity ligands; (g) optimal conditions for achieving improved product yield; and (h) optimal elution systems that minimize the denaturation of the purified antibodies.

Molecular modeling to rationalize ligand–support interactions in affinity chromatography

“…The downstream processing of protein-based pharmaceuticals represents in fact the main contemporary bottleneck for the large-scale production of this class of promising biomolecules. This technological challenge emerges clearly in the manufacturing of monoclonal antibodies (MABs), in which the costs associated with the state-of-the-art technology are due for a large part to the use of an affinity purification stage based on a natural ligand, Protein A [1,2]. The development of an effective and costefficient alternative to Protein A-based stationary phases has been the subject of much experimental and theoretical work [3][4][5][6][7][8][9][10].…”

Highly efficient and low-cost purification of lysozyme: A novel tris(hydroxymethyl)aminomethane immobilized affinity column

“…This may be due to the high cost of HPAC. The classical affinity ligands for the target protein, including antibodies [5,13,14], drugs [5], metal ions [15][16][17][18] and dyes [19][20][21], are either expensive or instable. The affinity columns synthesized with the corresponding affinity ligands are therefore high in cost and low in stability and repeatability.…”