Critical aspects on traditional antivenom production processes and their optimization by factorial analysis.

Critical aspects on traditional antivenom production processes and their optimization by factorial analysis.
Biologicals. 2020 Sep 07;:
Authors: Zurbano BN, Tavarone E, Viacava BG, Dokmetjian JC, Cascone O, Fingermann M
Abstract
Most antivenoms are produced by techniques developed over 50 years ago, with minor modifications. Herein we revise the core of traditional antivenom production processes aiming to optimize key determinants for both consistent antivenom production and the best balance between F(ab')2 quality and recovery. Factorial design analysis revealed that pepsin digestion of 1:3 saline diluted equine plasma for 60 min under pH: 3.20, 37 °C temperature and a 1:15 pepsin to protein ratio conditions, allowed to achieve maximal IgG to F(ab')2 conversion with minimal protein aggregate formation. Further downstream processing by salting out with ammonium sulfate was also studied by factorial analysis. The influence of ammonium sulfate (AS) concentration, temperature (T) and the albumin to total plasma protein ratio plasma (Alb:P) were assayed, revealing that both AS, T and their interaction have a significant impact in F(ab')2 quality and recovery. Taking into account the existing compromise between F(ab')2 monomer recovery and quality two alternative conditions were selected: 14 g/dl AS at 56 °C and, alternatively 16 g/dl AS at 30 °C. Reasonable yields (42%) and product quality (2.5% of aggregates) without significan...

Source: Biologicals : Journal of the International Association of Biological Standardization - September 6, 2020 Category: Biotechnology Authors: Zurbano BN, Tavarone E, Viacava BG, Dokmetjian JC, Cascone O, Fingermann M Tags: Biologicals Source Type: research