Comprehensive characterization of biotherapeutics is necessary to satisfy safety standards set by regulatory agencies and helps to ensure protein drug efficacy. Intact protein analysis using a combination of liquid chromatography (LC) and mass spectrometry (MS) provides information on the accurate mass of the protein and the relative abundance of its isoforms. The analysis of these complex heterogeneous protein biotherapeutics benefits from high resolution accurate mass (HRAM) instrumentation. Combining HRAM with powerful data deconvolution algorithms facilitates structural confirmation and accurate identification of protein modifications.

Intact mass analysis workflow

No need to de-glycosylate! Added information and confidence.

Using Orbitrap technology Drug to Antibody Ratio (DAR) analysis can be performed in native conditions, and without the need for deglycosylation. BioPharma Finder software provides automatic determination of DAR values. Performing antibody drug conjugate (ADC) analysis of the unaltered or native ADC mixture allows DAR value calculations to be done based on multiple glycoforms. In the example shown here the average DAR value is calculated using four different glycoforms and is very consistent, providing added information, DAR measurement accuracy and analytical confidence.

Resources

TypeTitle
Application Note Robust and Reproducible Peptide Mapping and Intact Mass Analysis Workflows on a Single Instrument Platform
Application Note SEC-MS with Volatile Buffers for Characterization of Biopharmaceuticals
Application Note Reversed-Phase Separation of Intact Therapeutic Antibodies Using the Vanquish Flex UHPLC System
Application Note Fast Online Desalting of mAbs Using a Reversed-Phase Desalting Cartridge for LC-MS Analysis
Intact Mass Methods AppsLab Library - Intact Mass Methods
Poster Note High Temperature, High Throughput Reversed Phase Separation of Intact Monoclonal Antibodies
Poster Note Enabling Mass Spectrometric Analysis of Intact Proteins in Native Conditions on A Hybrid Quadrupole-Orbitrap Mass Spectrometer
Poster Note Full Characterization of Heterogeneous Antibody Samples Under Denaturing and Native Conditions on a Hybrid Quadrupole-Orbitrap Mass Spectrometer
Poster Note Complete Characterization of a Cysteine-linked Antibody-Drug Conjugate Performed on a Hybrid Quadrupole-Orbitrap Mass Spectrometer with High Mass Range
Poster Note New Integrated Informatics Solution for Protein Biotherapeutics Characterization
Technical Note Influence of Column Temperature on Reversed-Phase Chromatography of an Intact Antibody

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