학술논문
Application of triple quadrupole mass spectrometry for the characterization of antibody-drug conjugates.
Document Type
Academic Journal
Author
Källsten M; Department of Chemistry-BMC, Analytical Chemistry, Uppsala University, Box 599, 751 24, Uppsala, Sweden. malin.kallsten@kemi.uu.se.; Recipharm OT Chemistry AB, Virdings allé 32b, 754 50, Uppsala, Sweden. malin.kallsten@kemi.uu.se.; Pijnappel M; Recipharm OT Chemistry AB, Virdings allé 32b, 754 50, Uppsala, Sweden.; Hartmann R; Department of Medicinal Chemistry, Organic Pharmaceutical Chemistry, Uppsala Biomedical Center, Uppsala University, Box 574, 751 23, Uppsala, Sweden.; Lehmann F; Oncopeptides AB, Luntmakargatan 46, SE-111 37, Stockholm, Sweden.; Kovac L; Recipharm OT Chemistry AB, Virdings allé 32b, 754 50, Uppsala, Sweden.; Lind SB; Department of Chemistry-BMC, Analytical Chemistry, Uppsala University, Box 599, 751 24, Uppsala, Sweden.; Bergquist J; Department of Chemistry-BMC, Analytical Chemistry, Uppsala University, Box 599, 751 24, Uppsala, Sweden. jonas.bergquist@kemi.uu.se.
Source
Publisher: Springer-Verlag Country of Publication: Germany NLM ID: 101134327 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1618-2650 (Electronic) Linking ISSN: 16182642 NLM ISO Abbreviation: Anal Bioanal Chem Subsets: MEDLINE
Subject
Language
English
Abstract
Antibody-drug conjugates (ADCs) are an inherently heterogeneous class of biotherapeutics, the development of which requires extensive characterization throughout. During the earliest phases of preclinical development, when synthetic routes towards the desired conjugate are being assessed, the main interest lies in the determination of the average drug-to-antibody ratio (DAR) of a given batch as well as information about different conjugation species. There has been a trend in mass spectrometry (MS)-based characterization of ADCs towards the use of high-resolving mass spectrometry for many of these analyses. Considering the high cost for such an instrument, the evaluation of cheaper and more accessible alternatives is highly motivated. We have therefore tested the applicability of a quadrupole mass analyzer for the aforementioned characterizations. Eight ADCs consisting of trastuzumab and varying stoichiometries of Mc-Val-Cit-PABC-monomethyl auristatin E conjugated to native cysteines were synthesized and served as test analytes. The average DAR value and molecular weights (Mw) of all detected chains from the quadrupole mass analyzer showed surprisingly high agreement with results obtained from a time-of-flight (TOF) mass analyzer and hydrophobic interaction chromatography (HIC)-derived values for all investigated ADC batches. Acquired Mw were within 80 ppm of TOF-derived values, and DAR was on average within 0.32 DAR units of HIC-derived values. Quadrupole mass spectrometers therefore represent a viable alternative for the characterization of ADC in early-stage development. Graphical abstract.