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Intact Protein and Subunit Analysis

Intact Protein and Subunit Analysis

Get the Resolution and Sensitivity You Need

To characterize therapeutic proteins, you need to have a fast and precise method to determine the molecular weight of intact proteins, key subunits, major modifications, and other impurities such as degradation products and fragments. Mass spectrometry has been widely used for protein analysis in the biopharmaceutical industry because it offers a wealth of information about the drug substance. For example, intact protein mass spectrometry can help to elucidate whether the correct protein has been expressed with the expected amino acid sequence. It can also offer insight on post-translational modifications by measuring the protein’s molecular weight accurately.

Mass spectrometry in conjunction with liquid chromatography can be a powerful purity solution. LC/MS techniques can provide information on relative abundances of different proteins, their isoforms, and impurities that are present in a sample. Protein analysis by LC/MS typically measures the intact proteins, deglycosylated proteins (for glycoproteins), antibody subunits such as reduced light chains and heavy chains, or IdeS digests to characterize the Fc and Fab fragments of the monoclonal antibody (mAb).

Agilent provides full workflow solutions, from sample preparation to data analysis, for the analysis of intact mAbs and key fragments. For the analysis of intact mAbs and antibody subunits, the Agilent AssayMAP Bravo platform brings reproducibility to a new level with high-precision liquid-handling robotics, and preprogrammed workflow templates that cover mAb purification, deglycosylation, reduction, and digestion with IdeS or other enzymes. Customized workflows are also possible, and the high-precision robotics improve speed and reproducibility for experiments of any scale.

The Agilent 6545XT AdvanceBio LC/Q-TOF system is designed for protein analysis with excellent mass accuracy, high resolution, high sensitivity, and wide dynamic range for simultaneous characterization of low- and high-abundance post-translational modifications. The Agilent 1290 Infinity II LC system provides high resolution, high performance protein separations.

What’s more, Agilent offers the most comprehensive range of optimized wide-pore reversed-phase columns for intact protein analysis. Our portfolio includes totally porous particles, superficially porous particles, and polymeric columns for analysis under the most extreme conditions. Agilent MassHunter BioConfirm software provides automatic data processing for MS deconvolution and protein sequence matching. Use it to streamline your analyses and organize the wealth of information that LC/MS offers.

Find on demand webinars and additional resources:  Intact Protein Analysis Resource Page

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