engleski

Influence of production conditions on IgG-based snake antivenoms’ stability properties

Antivenoms, having pure animal IgGs or their fragments as an active drug, are the only specific medicines against envenoming due to venomous animals’ bites. However, such products are of low sustainability for many reasons resulting in constant shortages all over the world. Stability of the product is one of them contributing not only to sustainability but it’s safety as well. It has been hypothesized that roughness of conditions to which IgGs are exposed during downstream purification disturbs more or less their conformation, making them consequently more prone to aggregation to varying degree, particularly after secondary stress exposure. The aim of this research was to investigate the impact of five commonly applied biochemical principles for IgG extraction from plasma on stability properties of pure IgGs. For that purpose, equine IgGs were purified from unique sample of hyperimmune plasma by two mild condition operational procedures (anion-exchange chromatography (AEX) and caprylic acid precipitation (CAP)) and three harsher ones (ammonium sulphate precipitation (ASP), cation- exchange chromatography (CEX) and affinity chromatography (AC)). Their stability was studied under non-optimal storage conditions (42 °C, transient lowering of pH) by monitoring the changes of aggregate content and thermal stability of pure IgG preparations. We found that gentle protocols initially generate IgGs with lower aggregate content in comparison to harsher ones. Their tendency for further aggregation was proportional to the initial aggregate share. Thermal stability of IgG molecules inversely correlated to the aggregate content in refined samples. We can conclude that mild condition purification protocols indeed generate more stable IgGs.

plasma processing ; IgG antivenom ; aggregates ; thermal stability

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