title: Development and scale-up of the recovery and purification of a domain antibody Fc fusion protein-comparison of a two and three-step approach.
abstract: A robust, economical process should leverage proven technology, yet be flexible enough to adopt emerging technologies which show significant benefit. Antibody and Fc-fusion processes may capitalize on the high selectivity of an affinity capture step by reducing the total number of chromatographic steps to 2. Risk associated with this approach stems from the potentially increased time frame needed for process development as well as unforeseen changes in impurity profile during first scale-up of drug substance (DS) for animal toxicology and clinical phase I trials (FIH) production, which could challenge a two-step process to the point of failure. Two different purification strategies were pursued during process development for an FIH process of a dAB-Fc fusion protein. A two-step process was compared to a three-step process. The two-step process leveraged additives to maximize impurity reduction during affinity capture. While wash additives in combination with a mixed mode chromatography met all impurity reduction requirements, HCP levels were still higher as compared to the three-step process. The three-step process was implemented for manufacture of clinical material to mitigate risk.
authors: Biologics Development, BLY-106G, Bristol-Myers Squibb, 519 Route 173 West, Bloomsbury, New Jersey. sibylle.herzer@bms.com.
authors: Sibylle
authors: S
authors: Herzer
authors: Biologics Development, BMS, Bloomsbury, New Jersey.
authors: Atul
authors: A
authors: Bhangale
authors: Biologics Development, BMS, Bloomsbury, New Jersey.
authors: Gregory
authors: G
authors: Barker
authors: Biologics Development, BMS, Bloomsbury, New Jersey.
authors: Isha
authors: I
authors: Chowdhary
authors: Biologics Development, BLY-106G, Bristol-Myers Squibb, 519 Route 173 West, Bloomsbury, New Jersey.
authors: Matthew
authors: M
authors: Conover
authors: Biologics Development, BMS, Seattle, Washington.
authors: Brian W
authors: BW
authors: O'Mara
authors: Biologics Development, BMS, Seattle, Washington.
authors: Lily
authors: L
authors: Tsang
authors: Gilead Sciences, Oceanside, California.
authors: Shi-Yu
authors: SY
authors: Wang
authors: CADD, BMS, Lawrenceville, New Jersey.
authors: Stanley R
authors: SR
authors: Krystek
authors: Biologics Development, BMS, Bloomsbury, New Jersey.
authors: Yan
authors: Y
authors: Yao
authors: Biologics Development, BMS, Bloomsbury, New Jersey.
authors: Siegfried
authors: S
authors: Rieble
mesh_terms: D000818:Animals; D016466:CHO Cells; D002853:Chromatography, Liquid / Q000379:methods*; D003412:Cricetulus; D007141:Immunoglobulin Fc Fragments / Q000235:genetics / Q000302:isolation & purification*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D007141:Immunoglobulin Fc Fragments; D011993:Recombinant Fusion Proteins
keywords: Fc-fusion protein; caprylate; design of experiments; process development; purification; scale-up