Overview of IVIG Process Development
The IVIG plasma fractionation process is a highly regulated, multi-step procedure designed to isolate and purify immunoglobulin G (IgG) from human plasma. The process begins with plasma collection from rigorously screened, healthy donors and employs the classic Cohn fractionation method—utilizing cold ethanol under controlled pH, temperature, and ionic strength—to separate plasma proteins. This method is complemented by subsequent purification steps, including chromatographic techniques and nanofiltration, to remove impurities and ensure viral inactivation. Analytical testing confirms that the final formulation achieves targeted concentration, pH, and osmolality, all of which are essential to enhance safety and tolerability during infusion.
Challenges of the Traditional Cohn Process
For over 70 years, the Cohn process has been the industry standard for IgG production. However, this method typically yields only 50–60% of the starting IgG and requires a 7–10 day production cycle. The process, which uses large volumes of ethanol combined with extreme pH changes, poses significant risks of protein denaturation, affecting stability and reducing the therapeutic efficacy of IVIG products. These factors also raise environmental concerns and highlight the need for improved methods that minimize these harsh conditions.
Learn more about the analytical data that reveals the limitations of the traditional Cohn process and why innovation is critical for patient safety
Addressing Procoagulant Contaminants
Procoagulant contaminants, such as plasma kallikrein, FXIIa, and FXIa, are a major concern in IVIG manufacturing because even trace amounts can trigger thrombotic events. Although adverse events occur in less than 1% of IVIG recipients, the potential for a hemostatic insult necessitates rigorous testing. Prolytix employs a suite of analytical methods—including NaPTT clot-based assays, PKA and FXIa chromogenic assays, high-resolution mass spectrometry (HRMS), and thrombin generation assays (TGA)—to detect and quantify these contaminants. These sensitive tests ensure that any activated coagulation factors are identified and removed, thereby mitigating the risk of adverse reactions.
Innovative Process Development with Prolytix
In response to the challenges posed by the traditional Cohn process, Prolytix developed a novel plasma fractionation technology that uses salt precipitation rather than ethanol. This innovative method reduces the process time to 48–72 hours and increases IgG yield to over 80%, compared to the conventional 50–60%. A critical enhancement in this process is the addition of an ion exchange “polishing” step. By spiking exogenous FXIa into the intermediate product and leveraging differences in ionic affinity via weak cation exchange, Prolytix was able to effectively remove FXIa contamination. This adaptation resulted in a final yield of approximately 77% and a purity of 99.7% pure IgG, as confirmed by HRMS and TGA tests, ensuring the product meets all European Pharmacopoeia (EP) safety metrics.
Reach out to learn more about our breakthrough analytical findings and process innovations that set new industry standards for safety and yield.
Collaboration and Process Validation Success
The journey to develop this next-generation IVIG manufacturing process was marked by a highly collaborative effort between Prolytix and its client over an 18-month period. During this time, weekly meetings and daily updates facilitated rapid troubleshooting and process refinement. Analytical data consistently guided the adjustments—ensuring contaminants were eliminated, as evidenced by undetectable FXIa levels via HRMS and TGA assays—and validated a scalable, eight-step process that not only shortens production timelines by three to six months but also delivers significant cost savings and enhanced patient safety.