Indian Journal of Veterinary Research (The)

  • Year: 2024
  • Volume: 33
  • Issue: 2

Protein G chromatography for an efficient purification of monoclonal antibodies

  • Author:
  • Samriddhi Dubey, Shreya Gopinath, Das Adwitiya, Katrapati Sai Lakshmikanth, Kailash Singh Bisht, Madhusudan Hosamani*
  • Total Page Count: 7
  • Published Online: Aug 27, 2025
  • Page Number: 7 to 13

ICAR-Indian Veterinary Research Institute, Hebbal, Bengaluru-560024, Karnataka

Abstract

This study presents an optimized Fast Protein Liquid Chromatography (FPLC)-based method for bench-scale purification of monoclonal antibodies (mAbs) using protein G affinity chromatography. Hybridoma clones 9C12F4 and 5G5H12 were cultured in serum-free medium, and their supernatants were applied for purification. Fine-tuned flow rates (0.5 mL/min binding, 1 mL/min elution) and precise pH control maximized recovery while preserving antibody functionality. The method yielded 1–2.8mg of Immunoglobulins (IgGs) per 100 mL culture supernatants for purifying IgG1 and IgG2b mAbs, ensuring efficient small-scale purification for reliable downstream applications.

Keywords

Affinity chromatography, Antibody purification, Monoclonal antibodies