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Development of sustainable antitoxin downstream processing (ANTI TOX NEW)

Project title: Development of sustainable antitoxin downstream processing (ANTI TOX NEW)

Grantor: Croatian Science Foundation

Grantor’s website:

Principal investigator: Beata Halassy, PhD

Duration: 15.09.2015.-14.02.2020.

Project’s web page:

Research team:   Tihana Kurtović, PhD

Maja Lang Balija, PhD

Julija Erhardt, PhD

Marija Brgles, PhD

Hrvoje Cajner, PhD

Monika Tunjić, mag.biol.mol.

Jasna Lalić, PhD

Dora Sviben, PhD

Sanja Mateljak Lukačević , MSc

Total funding: 992.000,00 HRK

Brief description:

Antitoxin is the general name for the antibodies of animal origin used to treat for example drug overdose (digoxin), bacterial toxins (diphtheria, tetanus and botulism), viral infections (rabies) and envenoming (snake, spider and scorpion). Antitoxins are produced by immunisation of animals, mostly horse and sheep, by appropriate antigen of interest (upstream part of the process), and subsequent purification of induced immunoglobulins from the hyperimmunised animal plasma (downstream part of the process). Established cost-effective antitoxin production has an additional value serving as a platform that can be easily adapted for fast production of therapeutics to combat new emerging diseases (ebola, for example).

The aim of the here proposed research is to design and optimise at the laboratory scale the most effective and most compact purification process (downstream part) based on the current scientific and technological knowledge. The process will be designed by balancing between fulfilling the growing regulatory demands, purification process yields (as high as possible) and the production costs (as low as possible). The aim is to find the optimal parameters of each purification step, to investigate the effectiveness of the designed process in reducing the risk of viral contamination, and to generate data on laboratory scale that could serve as the basis for decision making process regarding the sustainable antitoxin production plant establishment. The project will investigate for the first time the applicability of single-step simultaneous caprylic acid precipitation and peptic digestion of horse plasma, demonstrated so far only in ovine IgG purification, and the capability of this step in reducing the risk of viral inactivation, that has not been investigated yet. Further on, the applicability of monolithic chromatographic supports (convective interaction media) in the process of antitoxin purification will be explored for the first time.