title: Conformational analysis of recombinant monoclonal antibodies with hydrogen/deuterium exchange mass spectrometry.
abstract: Understanding the conformation of antibodies, especially those of therapeutic value, is of great interest. Many of the current analytical methods used to probe protein conformation face issues in the analysis of antibodies, either due to the nature of the antibody itself or due to the limitations of the method. One method that has recently been utilized for conformational analysis of antibodies is hydrogen/deuterium exchange mass spectrometry (H/DX MS). H/DX MS can be used to probe the conformation and dynamics of proteins in solution, requires small sample quantities, is compatible with many buffer systems, and provides peptide-level resolution. The application of H/DX MS to immunoglobulin gamma 1 (IgG1) recombinant monoclonal antibodies can provide information about IgG1 conformation, dynamics, and changes to conformation as a result of protein modification(s), changes in storage conditions, purification procedures, formulation, and many other parameters. In this article we provide a comprehensive H/DX MS protocol for the analysis of an antibody.
authors.affiliation: Biogen Idec, Inc., Cambridge, MA, USA.
authors.forename: Damian
authors.initials: D
authors.lastname: Houde
authors.forename: John R
authors.initials: JR
authors.lastname: Engen
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000302:isolation & purification; D002853:Chromatography, Liquid; D041961:Deuterium Exchange Measurement / Q000379:methods; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry* / Q000302:isolation & purification; D008970:Molecular Weight; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D012015:Reference Standards; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods / Q000592:standards; D053719:Tandem Mass Spectrometry / Q000379:methods / Q000592:standards
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D011994:Recombinant Proteins