Biotechnological production of drugs and fine chemicals

Course Description

COURSE CONTENT
1. Biotechnological drugs (biopharmaceuticals).
L: Biomanufacturing revolutions during history and representative products. Reference, generic, biosimilars and biobetters. Therapeutic proteins obtained by cell-free system technology. Glyco-engineering application in biopharmaceuticals improvement.
S: Challenges for biotechnological engineers in the 21 century regarding biopharmaceuticals.
2. Bioprocessing of biopharmaceutical production.
L: Upstream and downstream bioprocesses. Single-use technology application. Continuous bioprocessing. Modern, advanced and alternative separation techniques in biopharmaceutical production. Bioprocess integration. Final product formulation – Improvement of biopharmaceuticals’ stability and activity by current drug delivery technologies. Circular (recycling) economy in biopharmaceutical factory.
S: Next-generation biopharmaceutical production.
E: Industrial laboratory visit and exercises: Croatian Veterinary Institute Zagreb: application of Atomic absorption spectroscopy (AAS), Inductively coupled plasma atomic emission spectroscopy (ICP-OES) and Liquid chromatography–tandem mass spectrometry, Gas Chromatography - Tandem Mass Spectrometry (LC-MS-MS, GC-MS-MS) in conducting validated methods for biopharmaceuticals’ determination
3. Microbial biopharmaceuticals produced with natural and recombinant bacteria.
L: Production of biopharmaceuticals by E. coli - endotoxin detection and removal. Inclusion bodies separation - solubilization and renaturation of therapeutic proteins. „Live biotherapeutic products“ according to US FDA. Diabetes biopharmaceuticals, plasma proteins, recombinant proteins and microbial enzymes as biotherapeutics.
S: Production of biopharmaceuticals by bacteria.
E: Qualitative and quantitative determination of biopharmaceuticals by microbiological methods on solid and liquid culture media: detection of the microbial production in situ and determination of the unknown concentration of a pure substance by diffusion method.
4. Microbial biopharmaceuticals produced with natural and recombinant yeasts and moulds.
L: Biotechnological production of ergot alkaloids and fine biochemicals - exopolysaccharides, lipids, carotenoides.
S: Biopharmaceuticals production by yeasts and moulds.
E: Application of quantitative enzyme-linked immunosorbent assay (ELISA) for determination of biopharmaceutical residues
5. Antimicrobial biotherapeutics other than antibiotics.
L: Antiviral microbial drugs, enzybiotics, glycoconjugates, engineered lantibiotics, unnatural and beta-aminoacids therapeutic peptides, quorum-sensing inhibitors).
S: Biotechnological production of antimicrobial biotherapeutics other than antibiotics.
E: Detection of the genes involved in the biosynthesis of potential biotechnological drugs by PCR method
6. Biopharmaceuticals and fine chemicals of plant origin
L: Plants as a source of biopharmaceuticals and fine chemicals. Introduction in plant cell culture and obtaining of GM plants. Production of secondary metabolites, therapeutic proteins, and edible vaccines by plant.
S: Application of plant cell culture in biocatalytic processes in preparation of pharmaceuticals and fine chemicals
E: Introduction to plant cell culture: preparation of cell culture medium, callus induction and subculture of callus
7. Biopharmaceuticals of marine origin
P: Marine organisms as a source of biopharmaceuticals and fine chemicals. Introduction of algae cultivation and obtaining of GM algae. Production of biopharmaceuticals, therapeutic proteins, vaccines, and specific enzymes by algae. Production of algae for oral administration of biopharmaceuticals.
E: Isolation and identification of biologically active compounds from macroscopic algae
8. Animal cell cultures in biopharmaceuticals production
L: Animal cell cultures in biotechnology. Products of animal cell technology.
S: Recombinant proteins and vaccines produced by animal cell cultures.
E: Introduction to animal cell culture-cultivation, trypsinization and growth curve.
9. Stem cells and tissue engineering
L: Isolation of stem cells and therapeutic application. Bioengineering of tissue and tissue grafts.
S: Tissue engineering for therapeutic purposes.
10. Regulatory issues for biopharmaceuticals
L: Regulatory issues for research, production, clinical trials, application and marketing of biopharmaceuticals. Regulatory authorities (national, regional, international) HALMED, FDA, EMA, WHO. Patenting in biotechnology.
S: Patent documents of biopharmaceuticals.

LEARNING OUTCOMES

•     Compare biotechnological production of drugs by microbial, plant and animal cells and cell lines with the production of drugs via chemical synthesis, considering technological and environmental aspects.
• Justify the development of the biosimilar medicines and explain the application of contemporary engineering technologies, nanotechnology and microencapsulation aiming to achieve the drug with improved stability and activity
• Analyse process parameters in the biopharmaceuticals production to assess the efficacy of the biotechnological process
• Critically evaluate the pros and cons of batch versus continuous processes in biopharmaceutical production.
• Select the methods used for isolation and purification of the produced biotechnological drug, as well as methods used for the determination of active substance, potential contaminants and potential degradation products of the product.
• Explain the specifics of growing animal cell culture to produce tissues and organs
• Argue the benefits of using plant cell culture and single-use disposable biotechnological equipment for biopharmaceuticals production; perform a callus grafting procedure.
• Determine the concentration of biopharmaceuticals using microbiological and immuno-enzymatic ELISA methods and determine the presence of genes involved in the biosynthesis of potential biotechnological drugs by PCR method.
• Argue the importance of the continuous development of novel vaccines and modes of their application including the development of oral vaccines
• Explain the processes behind the development phases and approval of novel biopharmaceuticals

Literature

G. Jagschies, E. Lindskog, K. Lacki, P. M. Galliher (eds.) (2018) Biopharmaceutical processing: development, design, and implementation of manufacturing processes. Elsevier.

B. Kos, J. Novak, J. Šušković (2016) Mikrobni antagonizam i određivanje osjetljivosti mikroba na antimikrobne spojeve. In: Priručnik za vježbe iz opće mikrobiologije / Hajsig, D; Delaš, F. (eds.). Zagreb, Hrvatsko mikrobiološko društvo pp. 77-90.

J. Šušković, B. Kos, B. Šantek, Z. Kniewald, V. Mrša, M. Hruškar, J. Frece, V. Gaurina Srček, A. Slavica, J. Novak, A. Leboš Pavunc (2019) Biotehnologija u Hrvatskoj – povijesna baština i suvremeni trendovi, Annual of the Croatian Academy of Engineering, Vol. 2019 No. 1, pp. 438-484. file:///C:/Users/BLAENK~1/AppData/Local/Temp/Godisnjak_HATZ_2020_23__438_484.pdf

B. Kos, J. Šušković, J. Novak, A. Leboš Pavunc (2020) Bakteriocini i probiotici kao alternativne antimikrobne strategije u borbi protiv rezistencije na antibiotike. In: Antimikrobna rezistencija - izazovi i rješenja (I. Kosalec, ed.), textbook, University of Zagreb, pp. 186-211.

M.-K. Yeh (ed.) (2018) Biopharmaceuticals, IntechOpen, Open access peer-reviewed Edited Volume, https://www.intechopen.com/books/biopharmaceuticals

W. Jiskoot, G. F. A. Kersten, E. Mastrobattista, B. Slütter (2019) Vaccines. In: Crommelin D., Sindelar R., Meibohm B. (eds.) Pharmaceutical Biotechnology. Springer, Cham, pp. 281-304. https://link.springer.com/chapter/10.1007/978-3-030-00710-2_14#citeas

P. H. Rampelotto, A. Trincone (eds.) (2018) Grand Challenges in Marine Biotechnology, Springer (pdf format).

Z. Kniewald et al. (2000) Priručnik za pripravu i izolaciju biološki djelatnih supstancija, Alfej d.o.o., Zagreb.

Recent review and original scientific papers related to research of biotechnological production of drugs and fine chemicals.