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A novel method to clean protein G agarose for affinity column matrix with renewed binding capacity and high IgG selectivity.

Journal of immunological methods (2009-11-07)
Boaz O Owuor, Edmond J Remarque, Bart W Faber, Alan W Thomas
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.

MATERIALS
Product Number
Brand
Product Description

Millipore
Protein G–Agarose, lyophilized powder, Contains lactose stabilizers that must be removed prior to use.
Millipore
Protein G-Agarose, Fast Flow from Streptococcus sp., saline suspension