Vargas Mariángela, Segura Álvaro, Villalta Mauren, Herrera María, Gutiérrez José María, León Guillermo
Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
Biologicals. 2015 Jan;43(1):37-46. doi: 10.1016/j.biologicals.2014.10.003. Epub 2014 Nov 12.
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.
有必要在蛇毒抗毒素的生产中引入创新方法，以增加全球这些产品的供应。在这项工作中，将一种新方法（包括水两相系统（ATPS）作为主要纯化步骤）对马超免疫血浆的分级分离与传统的辛酸沉淀法进行了比较。用来自撒哈拉以南非洲的三种蛇的毒液免疫的马的超免疫血浆用作两种制剂生产的起始材料。在调整至相同的致死中和活性后，对两种抗毒素的免疫反应性、体外毒液活性的中和作用、理化特性和稳定性进行了比较。还评估了它们在产量和纯度方面的表现。通过ELISA和抗蛇毒血清学方法分析，两种制剂的体外酶活性中和谱和免疫反应性非常相似。同样，它们在稳定性研究中的表现也相似。然而，ATPS分级分离的抗毒素显示出改善的理化特性和免疫化学纯度及产量，主要是由于其较低的蛋白质含量。此外，该方法还能回收白蛋白作为副产品。ATPS纯化是一种有前途的抗毒素生产技术，应在临床前和临床水平上进一步评估。
