title: A reversed-phase high-performance liquid chromatography method for analysis of monoclonal antibody-maytansinoid immunoconjugates.
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.
authors.affiliation: ImmunoGen Inc., 128 Sidney Street, Cambridge, MA 02139, USA.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Fleming
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: John M
authors.initials: JM
authors.lastname: Lambert
authors.forename: Godfrey
authors.initials: G
authors.lastname: Amphlett
mesh_terms: D000911:Antibodies, Monoclonal / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D018796:Immunoconjugates / Q000032:analysis*; D008453:Maytansine / Q000031:analogs & derivatives* / Q000032:analysis; D015203:Reproducibility of Results
chemical_list: D000911:Antibodies, Monoclonal; D018796:Immunoconjugates; C488736:N(2')-deacetyl-N(2')-(3-mercapto-1-oxopropyl)maytansine; D008453:Maytansine