title: Bioreactor process parameter screening utilizing a Plackett-Burman design for a model monoclonal antibody.
abstract: Consistent high-quality antibody yield is a key goal for cell culture bioprocessing. This endpoint is typically achieved in commercial settings through product and process engineering of bioreactor parameters during development. When the process is complex and not optimized, small changes in composition and control may yield a finished product of less desirable quality. Therefore, changes proposed to currently validated processes usually require justification and are reported to the US FDA for approval. Recently, design-of-experiments-based approaches have been explored to rapidly and efficiently achieve this goal of optimized yield with a better understanding of product and process variables that affect a product's critical quality attributes. Here, we present a laboratory-scale model culture where we apply a Plackett-Burman screening design to parallel cultures to study the main effects of 11 process variables. This exercise allowed us to determine the relative importance of these variables and identify the most important factors to be further optimized in order to control both desirable and undesirable glycan profiles. We found engineering changes relating to culture temperature and nonessential amino acid supplementation significantly impacted glycan profiles associated with fucosylation, β-galactosylation, and sialylation. All of these are important for monoclonal antibody product quality.
authors.affiliation: Division of Product Quality Research, Office of Testing and Research, OPS, CDER, FDA, Silver Spring, Maryland.
authors.forename: Cyrus D
authors.initials: CD
authors.lastname: Agarabi
authors.affiliation: Biomolecular Measurement Division, Bioanalytical Science Group, National Institute of Standards and Technology, Gaithersburg, Maryland.
authors.forename: John E
authors.initials: JE
authors.lastname: Schiel
authors.affiliation: Division of Monoclonal Antibodies, Office of Biotechnology Products, OPS, CDER, FDA Silver Spring, Maryland.
authors.forename: Scott C
authors.initials: SC
authors.lastname: Lute
authors.affiliation: Division of Monoclonal Antibodies, Office of Biotechnology Products, OPS, CDER, FDA Silver Spring, Maryland.
authors.forename: Brittany K
authors.initials: BK
authors.lastname: Chavez
authors.affiliation: Division of Pharmaceutical Analysis, Office of Testing and Research, OPS, CDER, FDA, St. Louis, Missouri.
authors.forename: Michael T
authors.initials: MT
authors.lastname: Boyne
authors.affiliation: Division of Monoclonal Antibodies, Office of Biotechnology Products, OPS, CDER, FDA Silver Spring, Maryland.
authors.forename: Kurt A
authors.initials: KA
authors.lastname: Brorson
authors.affiliation: Division of Product Quality Research, Office of Testing and Research, OPS, CDER, FDA, Silver Spring, Maryland. Electronic address: Mansoor.Khan@fda.hhs.gov.
authors.forename: Mansoora
authors.initials: M
authors.lastname: Khan
authors.affiliation: Division of Monoclonal Antibodies, Office of Biotechnology Products, OPS, CDER, FDA Silver Spring, Maryland.
authors.forename: Erik K
authors.initials: EK
authors.lastname: Read
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000378:metabolism; D019149:Bioreactors; D002240:Carbohydrate Sequence; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D006031:Glycosylation; D006825:Hybridomas / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D007074:Immunoglobulin G / Q000737:chemistry* / Q000378:metabolism; D013058:Mass Spectrometry; D051379:Mice; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000737:chemistry* / Q000378:metabolism; D013696:Temperature
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D011134:Polysaccharides
keywords: Plackett-Burman; biotechnology; cell culture; design of experiments (DoE); glycan profiling; glycoprotein; glycosylation; mass spectrometry; monoclonal antibody; quality by design (QbD)