title: Application of high-resolution MS in the quantification of a therapeutic monoclonal antibody in human plasma.
abstract: BACKGROUND
  Monoclonal antibodies are the fastest growing class of protein therapeutics. Ligand-binding assays have been the technique of choice for the quantification of these large proteins; however, LC-MS and more recently LC-HRMS have been gaining momentum as robust alternatives for the bioanalysis of antibodies in biological matrices.
  
  
  RESULTS
  Optimization of sample preparation and LC-HRMS analysis in MRM(HR) mode has allowed us to develop a highly specific dual-peptide targeted assay for the quantification of Rituximab, in human plasma. The linearity of the assay was established from 1.0 to 200 µg/ml for both light and heavy chain surrogate peptides, with accuracy and precision within 15%.
  
  
  CONCLUSIONS
  LC-HRMS can be an effective tool for the quantification of monoclonal antibodies in regulated bioanalysis.
authors.affiliation: Algorithme Pharma Inc., 575 Armand-Frappier, Laval, Québec, Canada, H7V 4B3.
authors.forename: Kevork
authors.initials: K
authors.lastname: Mekhssian
authors.forename: Jean-Nicholas
authors.initials: JN
authors.lastname: Mess
authors.forename: Fabio
authors.initials: F
authors.lastname: Garofolo
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000097:blood* / Q000378:metabolism; D058846:Antibodies, Monoclonal, Murine-Derived / Q000097:blood / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry / Q000378:metabolism; D007147:Immunoglobulin Light Chains / Q000737:chemistry / Q000378:metabolism; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D010455:Peptides / Q000032:analysis; D000069283:Rituximab; D014357:Trypsin / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D058846:Antibodies, Monoclonal, Murine-Derived; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D010455:Peptides; D000069283:Rituximab; D014357:Trypsin