The adoption of LC-MS-based multi-attribute method (MAM) analysis and Multi-Angle Light Scattering (MALS) techniques for monitoring of biotherapeutic variation has progressed greatly over recent years. Directly assessing molecular attributes contributing to efficacy, safety, stability, and process robustness enables organizations to obtain more data without the ambiguity of traditional assays.
Often, this data is generated with greater sensitivity and dynamic range than legacy assays, complementing or eventually replacing these traditional analyses in development, manufacturing, and quality organizations. Waters systems, software, consumables, and automation support the analysis of multiple biopharmaceutical attributes, from characterization to routine monitoring.
Application Notebook: Multi-Attribute Methods for Biopharmaceutical Analysis
Enable LC-MS biopharmaceutical analyses in every GxP lab with Waters BioAccord LC-MS System, including high-resolution MS and waters_connect software for compliance-ready data analysis and reporting.
Accelerate multi attribute characterization and monitoring with the Xevo G3 QTof high-performance HRMS benchtop system, delivering accurate qualitative and quantitative results.
Eliminate the unpredictability of analyte losses due to metal interactions, which can be especially pronounced with biological molecules, using MaxPeak Premier columns and systems.
Gain vital assistance at every development stage with the DAWN MALS detector. Couple with Eclipse to measure binding affinity during discovery. Place downstream of any LC and perform advanced analysis of protein-conjugate ratio, oligomerization, and aggregation.
Directly monitor molecular weight, size, and impurities in real time with ultraDAWN that connects downstream of any purification and enrichment method, such as IEX, FPLC, or TFF.
Take single quadrupole performance to the next level. Monitor targeted attributes with the ACQUITY QDa II Mass Detector, which is easy to use and remarkably robust.
Bar graphs showing results presented in the Peptide MAM App for four monitored attributes, C-terminal Lysine, HT02 oxidation, HT07 oxidation, and HT03 deamidation. Injections are labeled as 1–3 for Remicade, 4–6 for Inflectra, 7–9 for Avsola, and 10–12 for Renflexis.
Peptide MAM App: New Peak Detection performed with defined criteria based on fold change, %base peak level, %isotopic similarity and retention time tolerance. Each peak passing the criteria (Right Side) is displayed for review. The peaks can be further examined using MS spectra to manually verify the NPD results.
Peptide MAM App: A monitored attribute peptide panel (%Abundance) is displayed for three modified (oxidized, deamidated, and glycosylated) peptides. Predefined pass/fail criteria based on user defined limits determine the color of bars, and the presence of a warning icon above them. The table, below, displays target criteria and results for the DTLMISR Ox attribute.
Selected product quality attributes for Remicade (infliximab) and three biosimilars subjected to thermal stress (all digested with RapiZyme Trypsin), T0, T1 (1 week), and T2 (2 weeks). Injections 1–3=Remicade (infliximab), 4–6=Inflectra (infliximab), 7–9=Avsola (infliximab), and 10–12=Renflexis (infliximab).
