engleski

Comparison of two downstream processes for efficient and sustainable antivenom preparation in terms of yield, final product purity and virus-inactivating potential

Snakebite envenoming has recently been recognized by a WHO as a highly relevant public health issue. The only validated treatment is immunotherapy with animal-derived antivenoms. Since these life-saving medications have been out of the mainstream of pharmaceutical development/manufacture for decades, number of drawbacks pertaining to their availability, safety and efficacy is becoming increasingly evident. One of the main reasons is associated with low sustainability of current productions and technological innovation is of great need. We have developed two highly efficient antivenom downstream processing strategies at the laboratory scale. Both involve caprylic acid fractionation of horse plasma for purification of IgGs, their pepsin-mediated digestion and final polishing by flow-through chromatography, but differ in number of processing steps which reflects on performance complexity. Here we provide comparison of two processes in terms of the final product quality. Both F(ab')2-based antivenoms were of high purity (100 and 97%), proving comparable to the purest currently available on the regulated market. As contaminants, minor traces of IgM, transthyretin and inter-alpha-trypsin inhibitor were identified. Each preparation was free from aggregates. Prevention of viral transmission as safety assuring requirement was assessed on mumps and measles vaccine strains. Their infectivity was reduced by more than 5 log during caprylic acid-involved processing steps. Additional inactivation was achieved by digestion step, which appeared more efficient for measles (4.6 log) than for mumps virus (2.6 log). High purity of the experimental antivenoms, aggregate absence and the virus-inactivating power of processing strategies, together with satisfactory yields, support their further development towards clinics.

Antivenom ; Plasma processing ; Caprylic acid ; Pepsin digestion ; Chromatography ; Virus inactivation

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