title: A workflow for absolute quantitation of large therapeutic proteins in biological samples at intact level using LC-HRMS.
abstract: OBJECTIVE
  The commonly used LC-MS workflow to quantify protein therapeutics in biological samples is 'bottom-up' approach. In this study, the aim is to establish 'top-down' approach for absolute quantitation of therapeutic antibodies or proteins of similar sizes in biological samples at intact level.
  
  
  METHODS
  Using a recombinant human monoclonal antibody as the model molecule, we present a workflow to measure large therapeutic proteins in plasma at intact level based on deconvoluted high-resolution MS (HRMS) peaks. A novel MultiQuant™ software function was developed to automatically deconvolute the peaks and process the data.
  
  
  CONCLUSIONS
  The workflow showed satisfying performance. This is a proof of concept study demonstrating the feasibility of bioanalysis of large therapeutic proteins at intact level using LC-HRMS.
authors.affiliation: Pharmacokinetics, Dynamics, & Metabolism (PDM), Janssen Research & Development, Johnson & Johnson, 1400 McKean Road, Spring House, PA 19477, USA.
authors.forename: Wenying
authors.initials: W
authors.lastname: Jian
authors.affiliation: Pharmacokinetics, Dynamics, & Metabolism (PDM), Janssen Research & Development, Johnson & Johnson, 1400 McKean Road, Spring House, PA 19477, USA.
authors.forename: Lijuan
authors.initials: L
authors.lastname: Kang
authors.affiliation: SCIEX, 71 Four Valley Drive, Concord, ON, Canada, L4K 4V8.
authors.forename: Lyle
authors.initials: L
authors.lastname: Burton
authors.affiliation: Pharmacokinetics, Dynamics, & Metabolism (PDM), Janssen Research & Development, Johnson & Johnson, 1400 McKean Road, Spring House, PA 19477, USA.
authors.forename: Naidong
authors.initials: N
authors.lastname: Weng
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D057230:Limit of Detection; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D011994:Recombinant Proteins / Q000032:analysis / Q000097:blood; D012984:Software; D057188:Workflow
chemical_list: D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins
keywords: LC-HRMS; bioanalysis; intact level; protein bioanalysis; quantitation