title: Bacteriophage and impurity carryover and total organic carbon release during extended protein A chromatography.
abstract: In the biopharmaceutical industry, column chromatography residuals are routinely assessed by the direct measurement of mock eluates. In this study, we evaluated virus and other impurity carryover between protein A cycles and the feasibility of using a total organic carbon (TOC) analyzer to monitor for column impurity leakage as a correlate for actual measured carryover in mock eluates. Commercial process intermediates were used in scaled down studies of two protein A media, ProSep A (Millipore, Bedford, MA, USA) and MabSelect SuRe (GE Healthcare, Uppsala, Sweden). The chromatography system was programmed to run up to 200 normal load/elution cycles with periodic blank cycles to measure protein and phage carryover, and water flush cycles to measure TOC release. Sustained phage carryover was evident in each study. Carryover and TOC release was lowest in the case where cleaning was most stringent (50 mM NaOH/0.5 M Na(2)SO(4) with MabSelect SuRe). The TOC analysis at this time does not appear to be a viable practical means of measuring impurity carryover; direct measurements in mock eluates appears to be more predictive of column performance.
authors.affiliation: Office of Biotechnology Products, Center for Drug Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Avenue, Silver Spring, MD 20993, USA.
authors.forename: Scott
authors.initials: S
authors.lastname: Lute
authors.forename: Kurt
authors.initials: K
authors.lastname: Brorson
mesh_terms: D000818:Animals; D001435:Bacteriophages / Q000737:chemistry*; D016466:CHO Cells; D002244:Carbon / Q000032:analysis*; D002846:Chromatography, Affinity / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D004364:Pharmaceutical Preparations / Q000032:analysis / Q000302:isolation & purification*; D011485:Protein Binding; D013205:Staphylococcal Protein A / Q000737:chemistry; D014764:Viral Proteins / Q000032:analysis / Q000302:isolation & purification
chemical_list: D004364:Pharmaceutical Preparations; D013205:Staphylococcal Protein A; D014764:Viral Proteins; D002244:Carbon