Purification of equine whole IgG snake antivenom by using an aqueous two phase system as a primary purification step

 

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Autores: Vargas Arroyo, Mariángela, Segura Ruiz, Álvaro, Villalta Arrieta, Mauren, Herrera Vega, María, Gutiérrez, José María, León Montero, Guillermo
Formato: artículo original
Fecha de Publicación:2015
Descripción:There is a need to introduce innovations in the manufacture of snake antivenoms to increase the supply of these products worldwide. In this work, the fractionation of equine hyperimmune plasma with a new methodology that includes an aqueous two phase system (ATPS) as a primary purification step was compared with the traditional method of caprylic acid precipitation. Hyperimmune plasma from horses immunized with the venoms of three snakes from sub-Saharan Africa was used as starting material for the production of both formulations. After being adjusted to the same lethal neutralizing activity, both antivenoms were compared in terms of their immunoreactivity, neutralization of in vitro venom activities, physicochemical characteristics, and stability. Their performance in terms of yield and purity was also assessed. The neutralization profile of in vitro enzymatic activities and the immunoreactivity, analyzed by ELISA and antivenomic approaches, were very similar for both preparations. Likewise, they behaved similarly in stability studies. However, ATPS-fractionated antivenom showed improved physicochemical profile and immunochemical purity and yield, mainly owing to its lower protein content. Additionally, this methodology allowed the recovery of albumin as a byproduct. ATPS purification constitutes a promising technology for antivenom production and should be further evaluated at preclinical and clinical levels.
País:Kérwá
Institución:Universidad de Costa Rica
Repositorio:Kérwá
OAI Identifier:oai:https://www.kerwa.ucr.ac.cr:10669/30340
Acceso en línea:http://www.sciencedirect.com/science/article/pii/S1045105614000992
https://hdl.handle.net/10669/30340
Access Level:acceso abierto
Palabra clave:Snake antivenom
Aqueous two phase system
Caprylic acid
Immunoglobulins
Snake venom