- 第3日 6月24日（金） P会場（501，502，503）
The increasing number and complexity of biotherapeutics are creating new challenges for biopharma analytical departments. While mass spectrometry offers many advantages in biopharmaceutical characterization, methods based on peptide mapping are labor intensive and time-consuming, generating large data sets requiring trained and skilled manual interpretation. A faster and less complex alternative to characterizing antibodies by peptide mapping is middle-up intact mass analysis, which utilizes novel enzymes that cleave antibodies at a highly specific site above or below the hinge, resulting in well-defined fragments (subunits) that are amenable to characterization by LC-MS. We present a middle-up intact mass analysis method that delivers comprehensive characterization of glycosylated and clipped species in therapeutic antibody samples using a conventional and a proprietary time-resolved deconvolution method.
Processing data with the conventional deconvolution method enabled detection of oxidized, glycosylated, and species corresponding to C-terminal lysine clipping and species where additional amino acids were absent at both the C- and N-termini of the antibody subunit chains. The time-resolved deconvolution algorithm enabled identification of additional low-abundance species, the plausibility of which could be easily evaluated by examining the two-dimensional ion map visualization in the software.