Enhancing monoclonal antibody stability during protein a chromatography using 2-methyl imidazolium dihydrogen phosphate.

Autor: Desai R; Department of Pharmaceutical Sciences and Technology, Institute of Chemical Technology, Mumbai 400019, India; Department of Biological Sciences and Biotechnology, Institute of Chemical Technology, Mumbai 400019, India., Jaiswal R; Department of Biological Sciences and Biotechnology, Institute of Chemical Technology, Mumbai 400019, India., Manchekar T; Department of Biological Sciences and Biotechnology, Institute of Chemical Technology, Mumbai 400019, India., Dugam S; Department of Pharmaceutical Sciences and Technology, Institute of Chemical Technology, Mumbai 400019, India., Jain R; Department of Biological Sciences and Biotechnology, Institute of Chemical Technology, Mumbai 400019, India. Electronic address: rd.jain@ictmumbai.edu.in., Dandekar P; Department of Pharmaceutical Sciences and Technology, Institute of Chemical Technology, Mumbai 400019, India. Electronic address: pd.jain@ictmumbai.edu.in.
Jazyk: angličtina
Zdroj: Journal of chromatography. A [J Chromatogr A] 2024 Sep 27; Vol. 1733, pp. 465263. Date of Electronic Publication: 2024 Aug 13.
DOI: 10.1016/j.chroma.2024.465263
Abstrakt: This study investigates the impact of 2-methyl imidazolium dihydrogen phosphate (2-MIDHP) on monoclonal antibody (mAb) aggregation during the Protein A purification stage, at a low pH (pH 3.0), and the viral inactivation phase. Size-exclusion high-performance liquid chromatography (SE-HPLC) and dynamic light scattering (DLS) were used to assess the mAb aggregation. Additionally, the influence of 2-MIDHP on mAb recovery, host cell protein (HCP) clearance, and Protein A leaching was investigated. Thermal stability of mAb, eluted in buffers containing 5 % to 25 % 2-MIDHP was analysed, using differential scanning calorimetry (DSC). Structural insights were obtained via circular dichroism (CD) and fluorescence spectroscopy. Our findings indicated that 2-MIDHP exerted a concentration-dependent protective effect against mAb aggregation, at the pH of 3.0. As the concentration of 2-MIDHP was increased from 0 % to 25 %, the aggregation was significantly reduced from 3.8 ± 0.01 % to 0.56 ± 0.002 %, as analysed by SE-HPLC. Addition of 2-MIDHP did not significantly impact the mAb recovery, HCP clearance, or Protein A leaching. DSC data supported these results, with higher 2-MIDHP concentrations leading to increased melting temperatures of mAb. CD and fluorescence spectroscopy revealed no significant changes in the secondary structure or aromatic residue environment in 2-MIDHP-treated samples, despite the observed reduction in aggregation. The results suggested that 2-MIDHP mitigated mAb aggregation during Protein A purification, possibly by stabilizing the protein structure under acidic stress conditions. These findings offer valuable insights for improving the robustness of mAb purification processes, enhancing product quality and yield.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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