title: Purification of antibodies by precipitating impurities using Polyethylene Glycol to enable a two chromatography step process.
abstract: The purification of antibodies by precipitating impurities using Polyethylene Glycol (PEG) was assessed with the objective of developing a two chromatography column purification process. A PEG precipitation method was evaluated for use in the industrial purification of recombinant monoclonal antibodies (MAbs). Effective and robust precipitation conditions including PEG concentration, pH, temperature, time, and protein concentration were identified for several different MAbs. A recovery process using two chromatography steps in combination with PEG precipitation gave acceptable yield and purity levels for IgG1 and IgG4 antibodies with a broad range of isoelectric points (pI). PEG precipitation removed host cell proteins (HCPs), high molecular weight species (HMWS), leached Protein A ligand, and host cell DNA to acceptable levels when run under appropriate conditions, and some endogenous virus removal was achieved.
authors.affiliation: Department of Purification Development, Process Technical Development, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA. Electronic address: giese.glen@gene.com.
authors.forename: Glen
authors.initials: G
authors.lastname: Giese
authors.forename: Adam
authors.initials: A
authors.lastname: Myrold
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Gorrell
authors.forename: Josefine
authors.initials: J
authors.lastname: Persson
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D016466:CHO Cells; D011232:Chemical Precipitation; D002846:Chromatography, Affinity / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods; D006224:Cricetinae; D003412:Cricetulus; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D011092:Polyethylene Glycols / Q000737:chemistry*; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D013696:Temperature; D014780:Viruses / Q000302:isolation & purification
chemical_list: D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D011092:Polyethylene Glycols
keywords: AIEX; Antibody; CIEX; CV; DOE; Design of Experiments; HCP; HMWS; HPLC; High Performance Liquid Chromatography; LRV; Log Reduction Value; MAb; MMV; Minute Mouse Virus; PEG; Polyethylene Glycol; Precipitation; Purification; SEC; anion exchange; cation exchange; column volume; high molecular weight species; host cell protein; monoclonal antibody; size exclusion chromatography