title: Characterization of Aggregation Propensity of a Human Fc-Fusion Protein Therapeutic by Hydrogen/Deuterium Exchange Mass Spectrometry.
abstract: Aggregation of protein therapeutics has long been a concern across different stages of manufacturing processes in the biopharmaceutical industry. It is often indicative of aberrant protein therapeutic higher-order structure. In this study, the aggregation propensity of a human Fc-fusion protein therapeutic was characterized. Hydrogen/deuterium exchange mass spectrometry (HDX-MS) was applied to examine the conformational dynamics of dimers collected from a bioreactor. HDX-MS data combined with spatial aggregation propensity calculations revealed a potential aggregation interface in the Fc domain. This study provides a general strategy for the characterization of the aggregation propensity of Fc-fusion proteins at the molecular level.Graphical Abstract.
authors.affiliation: Bioanalytical and Discovery Analytical Sciences, Research and Development, Bristol-Myers Squibb Company, Route 206 & Province Line Road, Princeton, NJ, 08543, USA.
authors.forename: Richard Y-C
authors.initials: RY
authors.lastname: Huang
authors.affiliation: Department of Chemistry and Chemical Biology, Northeastern University, 360 Huntington Avenue, Boston, MA, 02115, USA.
authors.forename: Roxana E
authors.initials: RE
authors.lastname: Iacob
authors.affiliation: Molecular Structure and Design, Research and Development, Bristol-Myers Squibb Company, Route 206 & Province Line Road, Princeton, NJ, 08543, USA.
authors.forename: Stanley R
authors.initials: SR
authors.lastname: Krystek
authors.affiliation: Biologics Development and Operations, Global Manufacturing & Supply, Bristol-Myers Squibb Company, 6000 Thompson Road, Syracuse, NY, 13057, USA.
authors.forename: Mi
authors.initials: M
authors.lastname: Jin
authors.affiliation: Biologics Development and Operations, Global Manufacturing & Supply, Bristol-Myers Squibb Company, 311 Pennington-Rocky Hill Road, Hopewell, NJ, 08534, USA.
authors.forename: Hui
authors.initials: H
authors.lastname: Wei
authors.affiliation: Biologics Development and Operations, Global Manufacturing & Supply, Bristol-Myers Squibb Company, 311 Pennington-Rocky Hill Road, Hopewell, NJ, 08534, USA.
authors.forename: Li
authors.initials: L
authors.lastname: Tao
authors.affiliation: Biologics Development and Operations, Global Manufacturing & Supply, Bristol-Myers Squibb Company, 311 Pennington-Rocky Hill Road, Hopewell, NJ, 08534, USA.
authors.forename: Tapan K
authors.initials: TK
authors.lastname: Das
authors.affiliation: Bioanalytical and Discovery Analytical Sciences, Research and Development, Bristol-Myers Squibb Company, Route 206 & Province Line Road, Princeton, NJ, 08543, USA.
authors.forename: Adrienne A
authors.initials: AA
authors.lastname: Tymiak
authors.affiliation: Department of Chemistry and Chemical Biology, Northeastern University, 360 Huntington Avenue, Boston, MA, 02115, USA.
authors.forename: John R
authors.initials: JR
authors.lastname: Engen
authors.affiliation: Bioanalytical and Discovery Analytical Sciences, Research and Development, Bristol-Myers Squibb Company, Route 206 & Province Line Road, Princeton, NJ, 08543, USA. guodong.chen@bms.com.
authors.forename: Guodong
authors.initials: G
authors.lastname: Chen
mesh_terms: D041961:Deuterium Exchange Measurement / Q000379:methods; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry*; D007074:Immunoglobulin G / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods*; D008958:Models, Molecular; D066329:Protein Aggregates; D011487:Protein Conformation; D055503:Protein Multimerization; D011993:Recombinant Fusion Proteins / Q000737:chemistry
chemical_list: D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D066329:Protein Aggregates; D011993:Recombinant Fusion Proteins
keywords: Aggregation propensity; Fc fusion protein; Hydrogen/Deuterium exchange mass spectrometry