Enhanced biotechnological process for antivenom production using Quality by Design methodology

Snakebites envenoming are a global public health problem that largely affects economically vulnerable populations. The primary recognized treatment involves hyperimmune serum, consisting of sterile and heterologous immunoglobulin solutions targeting specific antigens. One established production method in Vital Brazil Institute involves antibody digestion by pepsin, fractionation with ammonium sulfate salt, and thermocoagulation, yielding 30-40% and requiring a 5-day duration. This study aims to assess a novel production method centered on recovering the (Fab’)₂ fraction using caprylic acid, intending to implement a more efficient technological process. A new methodology was developed, defining digestion conditions (37 °C, pH 3.2; 2.5 g L^-1 of pepsin over 120 minutes at 150 rpm) and fractionation parameters (37 °C, pH 5.8; 2% caprylic acid over 60 minutes at 250 rpm). These conditions were established through the application of Quality by Design, supported by a critical risk analysis. This method reduced the usage of raw materials, process time, and labor cost per hour while maintaining yield with greater purity. These modifications were promising, resulting in higher purity and lower manufacturing costs, thereby expanding the prospect of achieving a more robust production process with a safer, optimized product. This allows meeting production goals and exploring new opportunities in foreign markets.