title: LC-MS quantification of protein drugs: validating protein LC-MS methods with predigestion immunocapture.
abstract: A refinement of protein LC-MS bioanalysis is to use predigestion immunoaffinity capture to extract the drug from matrix prior to digestion. Because of their increased sensitivity, such hybrid assays have been successfully validated and applied to a number of clinical studies; however, they can also be subject to potential interferences from antidrug antibodies, circulating ligands or other matrix components specific to patient populations and/or dosed subjects. The purpose of this paper is to describe validation experiments that measure immunocapture efficiency, digestion efficiency, matrix effect and selectivity/specificity that can be used during method optimization and validation to test the resistance of the method to these potential interferences. The designs and benefits of these experiments are discussed in this report using an actual assay case study.
authors.affiliation: Boehringer-Ingelheim Pharmaceuticals, Ridgefield, CT, USA.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Duggan
authors.affiliation: Boehringer-Ingelheim Pharmaceuticals, Ridgefield, CT, USA.
authors.forename: Bailuo
authors.initials: B
authors.lastname: Ren
authors.affiliation: Boehringer-Ingelheim Pharmaceuticals, Ridgefield, CT, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Mao
authors.affiliation: Boehringer-Ingelheim Pharmaceuticals, Ridgefield, CT, USA.
authors.forename: Lin-Zhi
authors.initials: LZ
authors.lastname: Chen
authors.affiliation: Boehringer-Ingelheim Pharmaceuticals, Ridgefield, CT, USA.
authors.forename: Elsy
authors.initials: E
authors.lastname: Philip
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry; D019921:Biotinylation; D005591:Chemical Fractionation / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D057230:Limit of Detection; D059346:Magnets / Q000737:chemistry; D004364:Pharmaceutical Preparations / Q000032:analysis* / Q000097:blood / Q000302:isolation & purification; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification; D059748:Proteolysis; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000911:Antibodies, Monoclonal; D004364:Pharmaceutical Preparations; D011506:Proteins
keywords: GXP bioanalysis; assay matrix effects; bioanalytical validation; digestion efficiency; hybrid bioanalysis; immunocapture; protein bioanalysis; protein drug validation; regulated bioanalysis; selectivity