engleski

The subtle change in the antivenom purification protocol affects differently intermediates and the final product

Animal-derived antibody therapeutics are widely used in medicine for viral and toxin neutralization. Although the various effective manufacturing methodologies have already been implemented into commercial scale production, due to more demanding regulatory requirements there is the constant need for their improvement. One of the major challenges in optimization concerns achieving the final product of high purity in order to ensure its safety and clinical efficiency. Recently, we have established the high yielding protocol for the preparation of snake venom- specific F(ab')2 fragments from hyperimmune horse plasma. The process itself consists of two main phases: caprylic acid selective precipitation of majority of contaminating proteins and pepsin fragmentation of IgG molecules preserved in solution. In the previous research, we have noticed that the most variable part of our refining scheme was caprylic acid fractionation step, thus affecting the purity of the IgG intermediate preparation. Despite the fact that we have succeeded to obtain the F(ab')2- based final product of 100% purity, we aimed for further process improvement regarding its stability by optimizing this critical step. Our goal was to investigate the possible beneficial impact of higher caprylic acid concentration on the IgG fraction composition in order to establish less variable and more reproducible manufacturing process. Here we showed the results indicating that even small variations in operating parameters could cumulatively affect refinement strategy for immunotherapeutics preparation, influencing consequently the quality and, particularly, safety of the final product.

antivenom ; purification ; caprylic acid

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