1. A reversed-phase high-performance liquid chromatography method for analysis of monoclonal antibody-maytansinoid immunoconjugates
- Author
-
Wei Zhang, Fleming Michael, John M. Lambert, and Godfrey W. Amphlett
- Subjects
musculoskeletal diseases ,Drug ,congenital, hereditary, and neonatal diseases and abnormalities ,Immunoconjugates ,medicine.drug_class ,media_common.quotation_subject ,Biophysics ,Maytansinoid ,Monoclonal antibody ,Biochemistry ,High-performance liquid chromatography ,chemistry.chemical_compound ,Ultraviolet visible spectroscopy ,medicine ,Maytansine ,Molecular Biology ,Chromatography, High Pressure Liquid ,media_common ,Chromatography ,Chemistry ,Antibodies, Monoclonal ,Reproducibility of Results ,Cell Biology ,Reversed-phase chromatography ,Selectivity ,Conjugate - Abstract
A reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed for detection and quantification of free maytansinoid drug in disulfide-linked conjugates between monoclonal antibodies and the maytansinoid drug DM1 (MAb-DM1). Mobile phases and gradient conditions were optimized for separation of several DM1-related free drug species from MAb-DM1 conjugates. The selectivity, linearity, and reproducibility of the method are reported. Reduction of the disulfide-linked DM1 followed by RP-HPLC allowed estimation of purity of MAb-linked DM1 as well as recovery of L-DM1. The method was also used to estimate drug per MAb ratios, which were consistent with those determined by UV spectroscopy.
- Published
- 2004