1R&D Division, Hetero Biopharma Limited, Jadcherla, Mahaboob Nagar, Telangana, India-509301
2Department of Biotechnology, Vignan's Foundation for Science Technology and Research (Deemed to be University), Guntur, Andhra Pradesh, India-522213
Therapeutic proteins such as monoclonal antibodies are complex molecules, need extensive characterization before their release into the market for clinical use. Therapeutic protein formulations are routinely characterized for their tertiary structure. Extrinsic fluorescence spectroscopy using various molecular rotors such as polarity sensitive ANS dyes is commonly used for the tertiary structure determination. However, the formulation excipient profile containing surfactants such polysorbate is known to have profound influence in terms of background noise in the extrinsic fluorescence when ANS dyes are used. However, there is neither any study that has clearly indicated the interference of polysorbate 80 in the extrinsic fluorescence profile of a formulated IgG1 monoclonal antibody drug product nor any study has reported the solution to the problem of avoiding interference from polysorbate 80 during extrinsic fluorescence analysis of a formulated monoclonal antibody drug product. Here in this study we reported the interference of polysorbate 80 in the extrinsic fluorescence profile of therapeutic protein drug product and recommending a polysorbate 80 removal step prior to extrinsic fluorescence analysis of drug product for the tertiary structure determination.
Polysorbate, extrinsic fluorescence, anti-CD20 antibody, tertiary structure, ANS dye
