Critical aspects on traditional antivenom production processes and their optimization by factorial analysis.
Biologicals
; 68: 65-73, 2020 Nov.
Article
en En
| MEDLINE
| ID: mdl-32912811
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 13 saline diluted equine plasma for 60 min under pH 3.20, 37 °C temperature and a 115 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 (AlbP) 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 significant changes in production cost of traditional methodologies was achieved under the optimized conditions found.
Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Mordeduras de Serpientes
/
Venenos de Serpiente
/
Fragmentos Fab de Inmunoglobulinas
/
Antivenenos
/
Pepsina A
/
Caballos
Límite:
Animals
/
Humans
Idioma:
En
Revista:
Biologicals
Asunto de la revista:
ALERGIA E IMUNOLOGIA
Año:
2020
Tipo del documento:
Article
País de afiliación:
Argentina