Enhanced biotechnological process for antivenom production using Quality by Design methodology

Abstract

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.

GRAPHICAL ABSTRACT

Keywords:

• Antivenom
• drug development
• process optimization
• Quality by Design
• technological innovation

Acknowledgements

The authors expres their thanks to the Conselho Nacional de Desenvolvimento Cientıfico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ – E-26/202.447/2019; FAPERJ – E-26/210.001/2021), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001 and Universidade Federal Fluminense and researcher Norman A. Ratcliffe.

Disclosure statement

No potential competing interest was reported by the authors.

Data availability statement

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Correction Statement

This article has been corrected with minor changes. These changes do not impact the academic content of the article.

Additional information

Funding

This work was supported by the Conselho Nacional de Desenvolvimento Cientıfico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ – E-26/202.447/2019; FAPERJ – E-26/210.001/2021), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001 and Universidade Federal Fluminense and researcher Norman A. Ratcliffe.