Justin Bueno - Deep-Ultraviolet Resonance Raman (DUVRR) Spectroscopy of Therapeutic Monoclonal Antibodies Subjected to Thermal Stress

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      Publication Details (including relevant citation   information):

    Anal. Chem., Article ASAP
    DOI: 10.1021/acs.analchem.5b01606
    Publication Date (Web): July 1, 2015


        The structural assessment of Rituximab, an IgG1 mAb, was   investigated with deep-ultraviolet resonance Raman (DUVRR)   spectroscopy. DUVRR spectroscopy was used to monitor the changes   to the secondary structure of Rituximab under thermal stress.   DUVRR spectra showed obvious changes from 22 to 72 °C.   Specifically, changes in the amide I vibrational mode were   assigned to an increase in unordered structure (random coil).   Structural changes in samples heated to 72 °C were related to   loss in drug potency via a complement dependent cytotoxicity   (CDC) bioassay. The DUVRR spectroscopic method shows promise as a   tool for the quality assessment of mAb drug products and would   represent an improvement over current methodology in terms of   analysis time and sample preparation. To determine the scope of   the method, protein pharmaceuticals of different molecular   weights (ranging from 4 to 143 kDa) and secondary structure   (β-sheet, α-helix and unordered structure) were analyzed. The   model illustrated the method’s sensitivity for the analysis of   protein drug products of different secondary structure. Results   show promise for DUVRR spectroscopy as a rapid screening tool of   a variety of formulated protein pharmaceuticals.

      Address (URL): http://pubs.acs.org/doi/full/10.1021/acs.analchem.5b01606