Herrera María, Solano Daniela, Gómez Aarón, Villalta Mauren, Vargas Mariángela, Sánchez Andrés, Gutiérrez José María, León Guillermo
Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica; Sección de Química Analítica, Escuela de Química, Universidad de Costa Rica, San José, Costa Rica.
Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
Toxicon. 2017 Feb;126:32-37. doi: 10.1016/j.toxicon.2016.12.004. Epub 2016 Dec 10.
Freeze-drying is a process used to improve the stability of pharmaceutical proteins, including snake antivenoms. This additional step confers these with a higher stability in comparison to liquid formulations, especially in tropical regions where high temperatures could affect the activity of immunoglobulins. Currently, the knowledge about freeze-drying process conditions for snake antivenoms is very limited. Some of the scarce scientific works on this subject reported reconstitution times up to 90 min for these preparations, which could imply a delay in the beginning of the antivenom therapy at the clinical setting. Therefore there is a reasonable concern about whether freeze-dried antivenoms exhibit the desired attributes for solid pharmaceutical proteins. In this work, a physicochemical characterization of seven commercial freeze-dried snake antivenoms was performed based on tests recommended by the World Health Organization (WHO). No significant differences were observed between the products regarding macroscopic appearance of the solid cakes, reconstitution times, residual humidity and monomers content. On the other hand, total protein concentration, turbidity and electrophoretic profile were different among samples. Microscopic analysis by scanning electron microscopy showed no collapsed structure and, instead, most of the samples showed a characteristic protein morphology composed of smooth plates and channels. All the parameters tested in this study were according to literature recommendations and evidenced that, in spite of slight variations found for some products, formulation and freeze-drying conditions chosen by manufacturers are adequate to prevent aggregation and generate, in physicochemical terms, freeze-dried antivenoms of acceptable quality.
冷冻干燥是一种用于提高包括蛇毒抗血清在内的药用蛋白质稳定性的工艺。与液体制剂相比，这一额外步骤赋予它们更高的稳定性，尤其是在高温可能影响免疫球蛋白活性的热带地区。目前，关于蛇毒抗血清冷冻干燥工艺条件的知识非常有限。关于这个主题的一些稀少的科学著作报道，这些制剂的复溶时间长达90分钟，这可能意味着在临床环境中抗蛇毒血清治疗开始的延迟。因此，人们合理地担心冷冻干燥的抗蛇毒血清是否具有固体药用蛋白质所需的特性。在这项工作中，根据世界卫生组织（WHO）推荐的测试方法，对七种市售冷冻干燥蛇毒抗血清进行了物理化学表征。在固体饼的宏观外观、复溶时间、残留湿度和单体含量方面，各产品之间未观察到显著差异。另一方面，样品之间的总蛋白浓度、浊度和电泳图谱有所不同。通过扫描电子显微镜进行的微观分析显示没有塌陷结构，相反，大多数样品显示出由光滑平板和通道组成的特征性蛋白质形态。本研究中测试的所有参数均符合文献建议，并且证明，尽管某些产品存在细微差异，但制造商选择的配方和冷冻干燥条件足以防止聚集，并在物理化学方面产生质量合格的冷冻干燥抗蛇毒血清。
