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

Ranjeet Desai1, Rahul Jaiswal2, Triveni Manchekar2

  • 1Department 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.

Journal of Chromatography. A
|August 18, 2024
PubMed

Insights

2-methyl imidazolium dihydrogen phosphate (2-MIDHP) significantly reduces monoclonal antibody (mAb) aggregation during Protein A purification at low pH. This additive enhances mAb stability without negatively impacting recovery or purity during bioprocessing.

Area of Science:

  • Biopharmaceutical Manufacturing
  • Protein Chemistry
  • Process Development

Background:

  • Monoclonal antibody (mAb) aggregation is a critical issue during downstream processing, potentially compromising product quality and efficacy.
  • Low pH conditions, often employed in Protein A chromatography and viral inactivation, can exacerbate mAb instability and promote aggregation.
  • Developing strategies to mitigate aggregation under these stressed conditions is crucial for robust biomanufacturing.

Purpose of the Study:

  • To investigate the impact of 2-methyl imidazolium dihydrogen phosphate (2-MIDHP) on mAb aggregation during Protein A purification at low pH.
  • To evaluate the effect of 2-MIDHP on mAb recovery, host cell protein (HCP) clearance, and Protein A leaching.
  • To assess the influence of 2-MIDHP on mAb thermal stability and structural integrity.

Main Methods:

  • Size-exclusion high-performance liquid chromatography (SE-HPLC) and dynamic light scattering (DLS) were used to quantify mAb aggregation.
  • Differential scanning calorimetry (DSC) was employed to analyze mAb thermal stability.
  • Circular dichroism (CD) and fluorescence spectroscopy were utilized to probe mAb structural changes.

Main Results:

  • 2-MIDHP demonstrated a concentration-dependent protective effect against mAb aggregation at pH 3.0, reducing aggregation from 3.8% to 0.56% with increasing concentrations.
  • No significant adverse effects were observed on mAb recovery, HCP clearance, or Protein A leaching.
  • DSC, CD, and fluorescence spectroscopy indicated that 2-MIDHP enhanced mAb thermal stability and maintained protein structure without causing denaturation.

Conclusions:

  • 2-MIDHP effectively mitigates mAb aggregation during low pH Protein A purification, likely by stabilizing the antibody structure.
  • The use of 2-MIDHP presents a promising strategy for improving the robustness and efficiency of mAb purification processes.
  • These findings contribute to enhancing the quality and yield of therapeutic monoclonal antibodies.
Keywords:
AggregationIonic liquidMonoclonal antibodyProtein A chromatographyPurification

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