title: A novel method to clean protein G agarose for affinity column matrix with renewed binding capacity and high IgG selectivity.
abstract: Attempts have been made at finding ways of cleaning used protein G agarose to revive their efficiency and make them reusable for purifying immunoglobulin G (IgG) in affinity column chromatography. A successful cleaning procedure that involved the use of 4-mol/L urea with 0.1-mol/L sodium hydroxide has been previously evaluated although it had some deleterious effect on the column binding capacity and compromised the resin efficiency. Efforts to develop base-tolerant affinity columns by using genetically engineered ligands such as the recombinant protein A from GE Healthcare have achieved some progress, with the column efficiency getting compromised to a lesser extent. However, genetically engineered ligands are even more expensive and may not be readily affordable in modest laboratory settings, especially if large-scale purchases are needed in routine use. We report here a novel and simple cleaning method involving the use of polyethylene glycol (PEG8000) that renews matrix-binding capacity comparable to a new resin while retaining high selectivity for IgG.
authors.affiliation: Department of Parasitology, Biomedical Primate Research Centre, Rijswijk, The Netherlands.
authors.forename: Boaz O
authors.initials: BO
authors.lastname: Owuor
authors.forename: Edmond J
authors.initials: EJ
authors.lastname: Remarque
authors.forename: Bart W
authors.initials: BW
authors.lastname: Faber
authors.forename: Alan W
authors.initials: AW
authors.lastname: Thomas
mesh_terms: D000915:Antibody Affinity; D001671:Biochemistry / Q000379:methods*; D002846:Chromatography, Affinity; D003902:Detergents; D006801:Humans; D007106:Immune Sera / Q000032:analysis; D007074:Immunoglobulin G / Q000378:metabolism; D011092:Polyethylene Glycols / Q000378:metabolism*; D011485:Protein Binding; D012117:Resins, Synthetic
chemical_list: D003902:Detergents; D007106:Immune Sera; D007074:Immunoglobulin G; D012117:Resins, Synthetic; D011092:Polyethylene Glycols