Naslov
Simple end effective approach for antivenom manufacturing

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

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

Izvornik
2017 Annual Meeting of the Croatian Immunological Society with EFIS on Tour Symposium / - Zagreb, 2017, 57-57

Skup
2017 Annual Meeting of the Croatian Immunological Society with EFIS on Tour Symposium

Mjesto i datum
Zagreb, Hrvatska, 20.10.2017. - 21.10.2017

Vrsta sudjelovanja
Poster

Vrsta recenzije
Domaća recenzija

Ključne riječi
antivenom ; plasma processing

Sažetak
INTRODUCTION: Antibody-based therapeutics play an important role in specific treatment of some medical emergencies (toxin and viral neutralisation, overcoming of immunoglobulin deficiencies). Their safety, effectiveness and availability are critically dependent on optimised, high yielding and low-cost manufacturing process. AIM: Here, we report simple, feasible and economically viable purification strategy for preparation of horse plasma-derived antivenom immunoglobulins and/or F(ab')2 fragments, that are irreplaceable for counteracting post-snakebite pathophysiological manifestations. MATERIALS AND METHODS: Fractionation of hyperimmune plasma pool was performed in only few simple and easily scalable purification steps, all designed to thoroughly keep desired IgG molecules or their fragments in solution, preventing possible degradation/aggregation of the active principle due to precipitation or binding to chromatographic supports. Firstly, unwanted plasma proteins, mostly albumin, were precipitated by caprylic acid. IgG-enriched fraction, refined from precipitating agent, was used for pepsin digestion. The final product, F(ab')2 fragments, was polished by ion-exchange chromatography on CIM QA disk under conditions which prefer binding of pepsin and by-products of enzymatic digestion exclusively. RESULTS AND CONCLUSION: Developed procedure gives completely pure and aggregates-free F(ab')2- based product of over 75% yield, as monitored by several in vitro methods. In addition, highly pure IgG fraction, an intermediate produced with almost no losses in the described procedure, is also in compliance with regulatory requirements concerning impurity content. So the described technology might serve as a platform for producing horse antitoxins of both types (Ig- and F(ab')2- based) and for other medical purposed, also.

Izvorni jezik
Engleski

Znanstvena područja
Biotehnologija

POVEZANOST RADA