Clinical Pharmacokinetic Assessment of an Anti-MAdCAM Monoclonal Antibody Therapeutic by LC-MS/MS

Abstract

Liquid chromatography tandem mass spectrometry (LC-MS/MS) has been shown to be a viable tool for preclinical pharmacokinetic (PK) analysis of monoclonal antibody (mAb) therapeutics. This work describes free and total PK assays for the mAb PF-00547,659 in serum of ulcerative colitis patients in a First-In-Human study [Vermeire, S. et al. Gut2011, 60 (8), 1068-1075]. The assay to measure free PF-00547,659 used immuno-enrichment with a biotinylated anti-idiotypic antibody and streptavidin magnetic beads. The total assay used enrichment by protein G magnetic beads. Following elution of PF-00547,659 from the beads, addition of an extended sequence stable isotope labeled peptide and trypsin digestion, a proteotypic peptide derived from the CDR region of the light chain of PF-00547,659 was quantified by LC-MS/MS. The free assay had a calibration range from 7.03 ng/mL to 450 ng/mL. The assay was precise and accurate with interbatch imprecision <16.5%, and interbatch inaccuracy <13.7% at all concentrations investigated during assay qualification. Results from LC-MS/MS methodologies are compared with historical immunoassay data originally acquired during the course of the clinical study. PK parameter estimates were highly correlated between the two analytical approaches. This work provides precedence that immunoaffinity LC-MS/MS can effectively be used to measure the serum concentrations of mAb therapeutics in clinical studies.

Quantification of a Proteotypic Peptide from Protein C Inhibitor by Liquid Chromatography–Free SISCAPA-MALDI Mass Spectrometry: Application to Identification of Recurrence of Prostate Cancer

“…However, validation of candidate biomarkers has lagged far behind their discovery, presenting a roadblock to the development of new diagnostic assays. Addressing the dilemma of biomarker validation for clinical use has been a purported goal in the field of proteomics over the past decade, yet in our informal literature surveys we could find only 3 publications in which specific proteins were measured by use of selected reaction-monitoring assays in more than 1000 patient samples (3)(4)(5).…”

“…To overcome the challenges inherent in using complex samples for selected reaction-monitoring assays (6 ), an immuno-mass spectrometry (MS) 5 assay known as stable isotope standards and capture by antipeptide antibodies (SISCAPA) was developed (7 ) in which a high-affinity antipeptide antibody was used to enrich peptides of interest from trypsin-digested human plasma or sera before MS analysis. Recently we reported a modified SISCAPA approach that significantly reduced background peptides, eliminating the need for upstream liquid chromatography (LC) (8 -9 ).…”

Quantification of active infliximab in human serum with liquid chromatography–tandem mass spectrometry using a tumor necrosis factor alpha -based pre-analytical sample purification and a stable isotopic labeled infliximab bio-similar as internal standard: A target-based, sensitive and cost-effective method

“…Albumin accounts for approximately 50% of total serum proteins, thus the removal of albumin from serum enhances the sensitivity of the method [16]. Other approaches focus on capturing IgGs in serum by means of immuno-affinity purification using either protein A, protein G, or anti human Fc antibodies attached onto a solid support such as magnetic beads [17][18][19][20][21][22][23]. Thereafter, the therapeutic antibody is eluted and digested using trypsin and finally one 'signature' peptide (a non-endogenous peptide) is selected for quantification.…”

LC–MS/MS Quantification of Next-Generation Biotherapeutics: A Case Study for an IgE Binding Nanobody in Cynomolgus Monkey Plasma

“…LC-MS/MS is also an alternative to traditional LBAs for bioanalysis of biotherapeutics, as has been demonstrated in both pre-clinical and clinical settings [4][5][7][8][9][10]. The approach is typically based on the analysis, via multiple reaction monitoring (MRM), of one or several proteotypic peptides, unique for the protein of interest, obtained following enzymatic digestion of the protein.…”

“…Due to the complex nature of samples, sample preparation is often a critical step to reach the required sensitivity. Chromatographic pre-fractionation, solid-phase extraction, protein depletion and immuno-affinity enrichment have successfully been applied to quantify low concentrations of proteins [7,[10][11][12][13][14][15][16][17][18][19].…”