Paper-PEG-based membranes for hydrophobic interaction chromatography: purification of monoclonal antibody.
This article discusses the preparation of novel Paper-PEG interpenetrating polymer network-based membranes as inexpensive alternative to currently available adsorptive membranes. The Paper-PEG membranes were developed for carrying out hydrophobic interaction membrane chromatography (HIMC). PEG is normally very hydrophilic but can undergo phase separation and become hydrophobic in the presence of high antichaotropic salt concentrations. Two variants of the Paper-PEG membranes, Paper-PEG 1 and Paper-PEG 2 were prepared by grafting different amounts of the polymer on filter paper and these were tested for their hydraulic properties and antibody binding capacity. The better of the two membranes (Paper-PEG 1) was then used for purifying the monoclonal antibody hIgG1-CD4 from simulated mammalian cell culture supernatant. The processing conditions required for purification were systematically optimized. The dynamic antibody binding capacity of the Paper-PEG 1 membrane was about 9 mg/mL of bed volume. A single step membrane chromatographic process using Paper-PEG 1 membrane gave high monoclonal antibody purity and recovery. The hydraulic permeability of the paper-based membrane was high and was maintained even after many runs, indicating that membrane fouling was negligible and the membrane was largely incompressible.
Yu
Deqiang
D
Department of Chemical Engineering, McMaster University, 1280 Main Street West, Hamilton, Ontario, Canada L8S 4L7.
Chen
Xiaonong
X
Pelton
Robert
R
Ghosh
Raja
R
D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods*; D002854:Chromatography, Paper / Q000379:methods*; D057927:Hydrophobic and Hydrophilic Interactions; D008567:Membranes, Artificial; D010209:Paper; D011092:Polyethylene Glycols / Q000737:chemistry*; D013205:Staphylococcal Protein A / Q000737:chemistry; D014462:Ultrafiltration / Q000295:instrumentation / Q000379:methods*
D000911:Antibodies, Monoclonal; D008567:Membranes, Artificial; D013205:Staphylococcal Protein A; D011092:Polyethylene Glycols