Liquid Chromatography-Mass Spectrometry (LC-MS) is a powerful analytical technique used for the precise determination of the molecular weight of protein s, peptides, and other biomolecules. By coupling liquid chromatography (LC) for separation with mass spectrometry (MS) for detection.
Key Advantages of LC-MS for Molecular Weight Determination:
- High Accuracy: LC-MS provides highly accurate molecular weight determination due to its high resolution and sensitivity.
- Structural Insights: In addition to molecular weight, LC-MS can reveal information about the structure of the molecule, such as modifications (e.g., glycosylation, phosphorylation), sequence, or post-translational modifications.
- Versatility: LC-MS is suitable for a wide range of biomolecules, including proteins, peptides, nucleic acids, and small molecules.
Quantification: LC-MS will accurately quantify the molecular weight of the sample.
For proteins such as monoclonal antibodies (mAbs), LC-MS is especially valuable for determining the intact and reduced molecular weight, identifying isoforms, and characterizing post-translational modifications. It also helps in confirming the presence of correct protein folding and heterogeneity in therapeutic proteins.
Aragen provides a highly sensitive and robust methods for estimation of protein molecular weight by using various methodology like; Intact (whole protein), Reduced (Light chain & Heavy chain) and Deglycosylated Intact (after removing the glycans from the protein).
Various mass species will be reported with less than 50ppm mass error to in-line with the regulatory expectations.
Reporting of the mass species as follows w.r.t various methodology: (see the Centroid and Deconvoluted Spectra).
Intact: G0F, G0F/G1F, G1F, G1F/G2F, G2F.
Reduced: Light chain (LC), Heay chain (HC)+G0F & Heavy chain (HC)+G1F
Deglycosylated Intact: Intact with K1/ Intact with K2
