Peptide mapping is crucial for the comprehensive characterization and monitoring of biopharmaceuticals. Obtaining adequate chromatographic resolution to reliably separate each digest fragment requires robust, reproducible methods with exceptional chromatographic performance.
Waters peptide mapping solutions combine automation, sample preparation, chromatographic separation, mass spectrometry, and overarching software workflows to deliver unparalleled analysis of complex protein and glycoprotein digests with high throughput. Waters qualitative and quantitative analysis solutions support effective characterization, simple method transfer, and compliance ready monitoring for routine laboratories. Meet your research and development goals with Waters rugged and robust peptide mapping workflows.
Peptide map characterization is essential for elucidating the primary amino acid structure and quality attributes of proteins, including monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs).
Following protein digestion, each subsequent peptide provides sequence information, retention time, and intensity to confirm the whole protein sequence. Waters provides powerful mass spectrometry hardware and software workflows to understand complex digest mixtures and develop methodologies fit for transfer to routine laboratories.
Peptide map characterization is essential for elucidating the primary amino acid structure and quality attributes of proteins, including monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs).
Following protein digestion, each subsequent peptide provides sequence information, retention time, and intensity to confirm the whole protein sequence. Waters provides powerful mass spectrometry hardware and software workflows to understand complex digest mixtures and develop methodologies fit for transfer to routine laboratories.
eBook: Peptide Mapping for Biotherapeutics
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.
Area (%) comparison of the ACQUITY UPLC I-Class PLUS System and ACQUITY UPLC H-Class PLUS Bio Binary System (with 340 µL mixer and gradient delay) over four injections. Data indicates that peak area is preserved between the systems.
The peptide mapping workflow of the BioAccord System provides PTM identifications. The figure shows the review panel following automated data processing. The component table contains: sequence information, m/z and neutral mass, mass tolerance, %rel. abundance as well as fragmentation information for each peptide. The summary table provides a concise view of selected peptide modifications across all samples. The consistency in retention times is shown in the bottom left panel using TIC’s for injections 1 and 25.
IC overlay of mAb Tryptic Digest Standard injections with IDC off and IDC enabled with thresholds of 5, 10, and 20. The associated table shows total file size, percent reduction, and data processing time with UNIFI Scientific Information System.
XIC profiles demonstrating the effect of particle surface chemistry (BEH and silica) on selectivities for selected NIST mAb tryptic peptides using 0.1% TFA in water/acetonitrile mobile phases. Chromatograms have been aligned based on the retention time of the H38. The MW and pI values for the peptides (Da) are provided in the table inset. Peak identities are provided in Table 3.
Relative abundance of missed and non-specific cleavages for NISTmAb digests prepared using the manual and automated PeptideWorks workflows and a leading immobilized trypsin kit. Error bars represent one standard deviation. PeptideWorks delivers a more complete and specific digestion.
1:5 enzyme:protein ratio digestions of Remicade, highlighting cleanliness of the baseline with RapiZyme trypsin (blue) vs. an industry leading competitor (black). Y-axis is % TIC intensity.
