title: Advance chromatin extraction enhances performance and productivity of cation exchange chromatography-based capture of Immunoglobulin G monoclonal antibodies.
abstract: The impact of host cell-derived chromatin was investigated on the performance and productivity of cation exchange chromatography as a method for capture-purification of an IgG monoclonal antibody. Cell culture supernatant was prepared for loading by titration to pH 6.0, dilution with water to a conductivity of 4mS/cm, then microfiltration to remove solids. DNA content was reduced 99% to 30ppm, histone host cell protein content by 76% to 6300ppm, non-histone host cell protein content by 15% to 321,000ppm, and aggregates from 33% to 15%. IgG recovery was 83%. An alternative preparation was performed, adding octanoic acid, allantoin, and electropositive particles to the harvest at pH 5.3, then removing solids. DNA content was reduced to<1 ppb, histones became undetectable, non-histones were reduced to 24,000ppm, and aggregates were reduced to 2.4%. IgG recovery was 95%. This treatment increased dynamic capacity (DBC) of cation exchange capture to 173g/L and enabled the column to reduce non-histone host proteins to 671ppm. Step recovery was 99%. A single multimodal polishing step further reduced them to 15ppm and aggregates to <0.1%. Overall process recovery was 89%. Productivity at feed stream IgG concentrations of 5-10g/L was roughly double the productivity of a same-size protein A column with a DBC of 55g/L.
authors.affiliation: Bioprocessing Technology Institute, 20 Biopolis Way, Centros #06-01, Singapore 138668, Singapore.
authors.forename: Rui
authors.initials: R
authors.lastname: Nian
authors.affiliation: Bioprocessing Technology Institute, 20 Biopolis Way, Centros #06-01, Singapore 138668, Singapore. Electronic address: pete_gagnon@mac.com.
authors.forename: Pete
authors.initials: P
authors.lastname: Gagnon
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D016466:CHO Cells; D002412:Cations; D002843:Chromatin / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D006657:Histones / Q000032:analysis; D007074:Immunoglobulin G / Q000302:isolation & purification*; D013205:Staphylococcal Protein A
chemical_list: D000911:Antibodies, Monoclonal; D002412:Cations; D002843:Chromatin; D006657:Histones; D007074:Immunoglobulin G; D013205:Staphylococcal Protein A
keywords: Capacity; Cation exchange capture; Chromatin; Contaminant reduction; IgG; Productivity