title: Quantitative bottom up analysis of infliximab in serum using protein A purification and integrated μLC-electrospray chip IonKey MS/MS technology.
abstract: BACKGROUND
  TNO Triskelion has applied its general workflow for the development of quantitative LC-MS methods for proteins in biological matrices to the quantification of infliximab in rat serum using bottom up μLC-MS/MS. Results/methodology: The general workflow consists of sample purification, analyte processing and LC-MS analysis. In the development of a quantitative μLC-MS/MS method for infliximab in rat serum the analyte processing part and the LC-MS part were optimized, in order to meet the different sample requirements of μLC-MS as compared with UPLC-MS. Using the optimized μLC-MS/MS method the LOQ was 75 ng/ml.
  
  
  CONCLUSIONS
  The present study showed that it is possible to gain sensitivity when going to smaller scale LC-MS (UPLC-MS to μLC-MS). Due to the combination of a modified sample preparation approach and the application of μLC-MS a lower LOQ could be achieved for infliximab compared with a previously developed UPLC-MS method.
authors.affiliation: TNO Triskelion, Utrechtseweg 48, 3704 HE Zeist, The Netherlands.
authors.forename: Anne J
authors.initials: AJ
authors.lastname: Kleinnijenhuis
authors.affiliation: TNO Triskelion, Utrechtseweg 48, 3704 HE Zeist, The Netherlands.
authors.forename: Martha
authors.initials: M
authors.lastname: Ingola
authors.affiliation: TNO Triskelion, Utrechtseweg 48, 3704 HE Zeist, The Netherlands.
authors.forename: Jan H
authors.initials: JH
authors.lastname: Toersche
authors.affiliation: TNO Triskelion, Utrechtseweg 48, 3704 HE Zeist, The Netherlands.
authors.forename: Frédérique L
authors.initials: FL
authors.lastname: van Holthoon
authors.affiliation: TNO Triskelion, Utrechtseweg 48, 3704 HE Zeist, The Netherlands.|ProQR Therapeutics, Darwinweg 24, 2333 CR Leiden, The Netherlands.
authors.forename: William D
authors.initials: WD
authors.lastname: van Dongen
mesh_terms: D000818:Animals; D018501:Antirheumatic Agents / Q000097:blood*; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D000069285:Infliximab / Q000097:blood*; D056656:Lab-On-A-Chip Devices; D057230:Limit of Detection; D051381:Rats; D018401:Sample Size; D013205:Staphylococcal Protein A / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000295:instrumentation / Q000379:methods
chemical_list: D018501:Antirheumatic Agents; D013205:Staphylococcal Protein A; D000069285:Infliximab
keywords: absolute recovery; analyte processing; infliximab; protein A; μLC–MS/MS