May 7, 2019 – MSDG Meeting

The success of any assay is heavily dependent on the reagents used. For most assays this involves reagent antibodies, but in the case of biomarker assays recombinant proteins that act as a reference material for method development and QC are reagents that must be generated and characterized. Ensuring the quality of this reference protein is critical to the assay development, however oftentimes the only QC performed on these calibrator proteins is gel shift assay, which while useful only confirms if a protein is relatively close to the correct expected molecular weight. In order to provide a more accurate results, our group has utilized intact mass analysis to confirm amino acid sequences of calibrator proteins allowing for better quality control (QC) of these regents. In addition to this we have recently started to apply bottom-up proteomics techniques to our biomarker analysis in an effort to get additional information about these calibrator proteins. In this talk, two examples of how we are leveraging bottom-up to improve our reagent biomarkers workflows will be highlighted. The first example will explain how we are using proteomics to obtain sequence information for calibrator proteins that have amino acid sequences that are uncertain. For the second example our ongoing efforts to couple proteomics with immunocapture in order to characterize endogenous biomarkers will be discussed.

Mass spectrometry (MS) has long been an enabling technology in the regulatory definition of a well‐ characterized biotherapeutic protein.  It is an invaluable tool to characterize molecular structures, assess process variabilities, and monitor the critical quality attributes of biotherapeutics – but until just recently has it been made accessible to biochemists.  Recent advances in LC, MS and informatics technologies are transforming biotherapeutic analyses, not only in terms of the quality of analytical data, but also in their accessibility for biochemists to generate, capture, and harness top‐quality LC/MS information to enable faster decision making.This talk will focus on how the evolving requirements of biotherapeutic analysis are addressed using modern LCMS analytical workflows built upon the recent introduced BioAccord system, and on how the leading separation, spectrometric, and informatics technologies are expanding the range of analytical challenges that MS workflows can routinely address while maintaining data integrity. We will provide several case studies on a panel of monoclonal antibodies (mAbs) for routine characterization and product quality attributes monitoring, and highlight how a workflow‐driven approach can produce time‐of‐flight mass spectrometry data that is not only highly comparable in quality to more complex advanced MS instruments, but that is more available to non‐traditional MS users in biopharmaceutical development and manufacturing organizations.