Purification of Recombinant Proteins on NuviaTM cPrimeTM Hydrophobic Cation Exchange Media : A Simple Approach to Method Development

General Bovine serum albumin, bovine carbonic anhydrase, and conalbumin were purchased from Sigma-Aldrich. Lactoferrin was obtained from Glanbia Nutritionals, Inc. Monoclonal antibody, mAbX, was overproduced in a Chinese hamster ovary (CHO) cell culture and previously purified by column chromatographic methods. Protein fractions were analyzed by SDS-PAGE using Criterion Tris-HCl 4–20% linear gradient gels (Bio-Rad) stained with Bio-Safe Coomassie stain (Bio-Rad), and quantified on a GS-800 calibrated densitometer (Bio-Rad). The clearance of E. coli host cell proteins (HCPs) and double-stranded DNA (dsDNA) were determined by E. coli HCP ELISA kit F410 (Cygnus Technologies) and Quant-iT dsDNA High-Sensitivity Assay Kit (Invitrogen), respectively. Protein concentration was determined by UV absorption at 280 nm, using the respective coefficients at 1 mg/ml.