title: A Size-Exclusion Chromatography Method for Analysis of Clostridium difficile Vaccine Toxins.
abstract: High-performance size-exclusion chromatography (HPSEC or SEC) is a method that can be applied to measure size distribution of proteins, including aggregates, monomers, and fragments. In the biopharmaceutical industry the quantitation of aggregates contained in biotherapeutics and protein-based vaccines is critical given the potential impact on safety, immunogenicity, and efficacy. Hence, aggregation analysis of therapeutic proteins or protein-based vaccine products is almost always a requirement of regulatory agencies. SEC, also referred to as gel-filtration chromatography, separates molecules by size through a porous resin stationary phase. Under isocratic flow small molecules are retained on the column longer than large molecules. Here we describe the use of this SEC technique to characterize aggregation levels for four different protein antigens for a Clostridium difficile vaccine.
authors: Vaccine Analytical Department, Merck Research Laboratories, Merck & Co., Inc., 770 Sumneytown Pike, P.O. Box 4, WP42A-30, West point, PA, 19486, USA. catherine_lancaster@merck.com.
authors: Catherine
authors: C
authors: Lancaster
authors: Vaccine Analytical Department, Merck Research Laboratories, Merck & Co., Inc., 770 Sumneytown Pike, P.O. Box 4, WP42A-30, West point, PA, 19486, USA.
authors: Richard R
authors: RR
authors: Rustandi
authors: Merck & Co., Inc., West point, PA, USA.|Johnson & Johnson, Spring House, PA, USA.
authors: Paola
authors: P
authors: Pannizzo
authors: Vaccine Analytical Department, Merck Research Laboratories, Merck & Co., Inc., 770 Sumneytown Pike, P.O. Box 4, WP42A-30, West point, PA, 19486, USA.
authors: Sha
authors: S
authors: Ha
mesh_terms: D036002:ADP Ribose Transferases / Q000737:chemistry / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification; D001427:Bacterial Toxins / Q000737:chemistry / Q000302:isolation & purification; D001428:Bacterial Vaccines / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D002850:Chromatography, Gel / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D016360:Clostridioides difficile / Q000737:chemistry / Q000276:immunology*; D004761:Enterocolitis, Pseudomembranous / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control*; D004768:Enterotoxins / Q000737:chemistry / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005557:Formaldehyde / Q000737:chemistry; D006801:Humans; D066329:Protein Aggregates; D064166:Vaccine Potency; D014613:Vaccines, Attenuated; D014614:Vaccines, Synthetic
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; D001428:Bacterial Vaccines; D004768:Enterotoxins; D066329:Protein Aggregates; D014613:Vaccines, Attenuated; D014614:Vaccines, Synthetic; C041348:tcdA protein, Clostridium difficile; C031815:toxB protein, Clostridium difficile; D005557:Formaldehyde; D036002:ADP Ribose Transferases; C410164:actin-specific ADP-ribosyltransferase, Clostridium
keywords: Binary toxin; C. difficile; HPLC; Size-exclusion chromatography; TcdA; TcdB; Vaccine