Development of a novel manufacturing process for highly purified and stable immunoglobulin M with enhanced plasma utilization

BACKGROUND AND OBJECTIVES: Immunoglobulin M (IgM) is a promising therapeutic agent. However, its production is limited by high molecular weight, complex structure and poor stability. This study aimed to develop a novel manufacturing process for high-purity, stable plasma-derived IgM while improving plasma utilization. MATERIALS AND METHODS: The flow-through fraction from first-step anion-exchange chromatography (Step 1 AEX) in an intravenous immunoglobulin manufacturing process was used as the starting material. Second-step AEX (Step 2 AEX) and third-step mixed-mode chromatography (Step 3 MMC) were used to remove impurities, including prekallikrein activator (PKA), immunoglobulin G (IgG), immunoglobulin A (IgA) and α2-macroglobulin. The elution conditions for Step 2 AEX were optimized to improve IgM recovery and purity. Four resins were evaluated for Step 3 MMC, and separation conditions were optimized to increase IgM purity and recovery. Finally, formulation studies were conducted to improve IgM stability. RESULTS: The optimized conditions were stepwise elution for Step 2 AEX (0.4 M followed by 1.0 M sodium acetate), Capto Core 400 resin for flow-through-mode Step 3 MMC and loading at pH 7.8, conductivity 15.0 mS/cm and protein concentration 1.0% (w/v). Key impurities IgA and PKA were reduced to 0.184 ± 0.011 mg/mL and below the limit of detection (<10 IU/mL), respectively, and IgM purity exceeded 90%. Under the final formulation conditions (0.1 M glycine, pH 4.0), the high-purity IgM showed good stability. CONCLUSION: A novel process was developed to address the critical challenges in plasma-derived IgM manufacture, achieving high IgM purity (>90%). By optimizing plasma utilization, this process represents a promising platform for next-generation immunoglobulin therapies.