| Module Description: the focus of the module will be on the downstream aspects of biotechnology production processes, spanning the range of biomolecule purification and characterization techniques commonly deployed across the biopharmaceutical and food biotech sectors, and also dealing with the applications of enzymes as a specific product class.
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Lecture programme
Downstream processing methods used to recover and purify biomolecules at industrial scale. Release of intracellular proteins: physical, chemical and biological methods. Primary clarification methods; centrifugation and filtration. Concentration and primary purification: ultrafiltration and precipitation. Protein purification by chromatography: gel filtration, ion-exchange, affinity, hydrophobic interaction and hydroxyapatite methods. Liquid and dried formulations (freeze-drying and spray-drying).
Impurities potentially present: microbial, viral, pyrogenic, DNA, protein, chemical contaminants. Excipients and protein stabilization. Manufacturing criteria for different product classes.
Enzyme technology. Production, purification and application of: i. bulk enzymes (carbohydrases, proteases, lipases, cellulases, pectinases); ii therapeutic enzymes (asparaginase, DNase, glucocerebrosidase); iii. Diagnostic enzymes (glucose oxidase, β-galactosidase, alkaline phosphatase). The use of enzymes in industrial biotransformations (eg., penicillin acylase, aminoacylase, glucose isomerase). Types of immobilization methods and systems (enzymes and whole cells).
Practical programme:
Downstream processing
Downstream processing methods used to recover and purify biomolecules at industrial scale. Release of intracellular proteins: physical, chemical and biological methods. Primary clarification methods; centrifugation and filtration. Concentration and primary purification: ultrafiltration and precipitation. Protein purification by chromatography: gel filtration, ion-exchange, affinity, hydrophobic interaction and hydroxyapatite methods. Liquid and dried formulations (freeze-drying and spray-drying).Impurities potentially present: microbial, viral, pyrogenic, DNA, protein, chemical contaminants. Excipients and protein stabilization. Manufacturing criteria for different product classes.
Enzyme Technology
Enzyme technology. Production, purification and application of: i. bulk enzymes (carbohydrases, proteases, lipases, cellulases, pectinases); ii therapeutic enzymes (asparaginase, DNase, glucocerebrosidase); iii. Diagnostic enzymes (glucose oxidase, β-galactosidase, alkaline phosphatase). The use of enzymes in industrial biotransformations (eg., penicillin acylase, aminoacylase, glucose isomerase). Types of immobilization methods and systems (enzymes and whole cells).
Practical programme
Aspergillus oryzae fermentation and extracellular amylase production: A. oryzae fermentation monitoring dry cell weight, α-amylase activity, protease activity, soluble protein and pH. α-amylase recovery and primary purification – centrifugation and ultrafiltration, ion exchange, hydrophobic interaction and size exclusion chromatography. Product characterization via SDS-PAGE.
Learning outcomes will be achieved through a combination of lectures, practicals, problem solving exercises and self-directed learning
| Module Content & Assessment | |
|---|---|
| Assessment Breakdown | % |
| Formal Examination | 60 |
| Other Assessment(s) | 40 |