Naslov
A compact and effective procedure for antivenom downstream processing

Autori
Tihana Kurtovic, Maja Lang Balija, Monika Tunjić, Marija Brgles, Beata Halassy

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
4th International Symposium Venoms 2017 Book / Rowan, Edward G. - Oxford, 2017, 38-38

Skup
4th International Symposium Venoms 2017

Mjesto i datum
Oxford, Ujedinjeno Kraljevstvo, 29.08.2017. - 30.08.2017

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
antivenom production ; optimisation of manufacturing process ; hyperimmune equine plasma

Sažetak
Antivenoms obtained from hyperimmune animal plasma, mostly equine or ovine, are the only specific therapeutics effective for rapidly counteracting post-snakebite pathophysiological manifestations. There are various well established refinement processing strategies that have been implemented into commercial scale antivenom production. However, optimisation of manufacturing process yielding safe, effective and available immunotherapeutics is still of great concern. Here, we report simple, feasible and economically viable fractionation protocol for preparation of V. a. ammodytes-specific antivenom. Hyperimmune equine plasma pool was fractionated in only few simple and easily scalable purification steps. The purification process can be stopped at different steps, depending on the desired final antivenom product: whole IgG molecules or F(ab')2 fragments, both fulfilling regulatory requirements for product purity. Particular emphasis was put on quantification of each purification step in terms of IgG or F(ab')2 yield determined by several in vitro methods. Throughout the procedure, IgG molecules or F(ab')2 fragments were constantly kept in the solution, preventing their precipitation and binding to chromatography columns which can lead to aggregation. Firstly, unwanted plasma proteins, mostly albumin, were precipitated by caprylic acid (CA). Ultrafiltered IgG-rich and CAdepleted supernatant was used for pepsin digestion. The final product, F(ab')2 fragments, was polished by ionexchange chromatography on CIM QA disk under conditions which prefer binding of pepsin and byproducts of enzymatic digestion exclusively.

Izvorni jezik
Engleski

Znanstvena područja
Kemija, Biologija, Biotehnologija

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