Class Protein Production and Manipulation

This curricular unit is designed to provide students with a comprehensive understanding of current techniques used in the production and manipulation of proteins, which are essential in various fields, including research laboratories, biotechnology, the pharmaceutical industry or medicine.

Not applicable

1 Introduction to protein production. Proteins used in medicine and industry. 2. Proteins: function and structure. Challenges in protein production. 3. Production of recombinant proteins. Different hosts. Advantages and challenges. 3.1 Protein production in prokaryotes. Cloning strategies. 3.2 Expression vectors-promoters, replication origins, selection markers, tags. 3.3 Synthetic biology and reprogramming of cellular expression machineries. 3.4 New strategies to overcome the challenges of protein production in bacteria 3.5 Protein production in eukaryotic cells. 4. Protein engineering. Targeted and random mutagenesis - methods and applications. Genetically modified proteins: functional improvement and chimerogenesis. 5. protein purification. 5.1 Design of purification strategies. 5.2 Low- and high-resolution protein purification methods. 5.3 Methods for assessing the purity, structural integrity, and stability of a protein. 6. Study cases.

The lectures will be expository, supported by discussions of case studies. In the case study discussions, the different strategies followed for different proteins will be discussed with the students. Diverse cases will be provided in which different cloning, expression and purification strategies were chosen and these options will be discussed with the students. In this CU the students will carry out part of their learning through problem-solving learning. This course has a strong practical component in which students will carry out experimental work throughout the semester, in which they will clone, express and purify a protein, ultimately assessing its purity and stability. In parallel, students will carry out targeted mutagenesis of this protein. Students will take an active part in choosing the cloning strategy, purifying and assessing the stability and purity of the purified protein.

 Protein Downstream Processing-Design (Development, and Application of High and Low-Resolution Methods) - Methods in Molecular Biology-Springer protocols , Ed. Nikolaos E. Labrou, 2021, Human Press. Methods in Recombinant Protein Production, Editors: Volker Gurtler, Peter Smooker, Marek Kieliszek 1st Ed, 2022, Academic Press. Serão também disponibilizados artigos recentes sobre as diferentes técnicas e casos de estudo discutidos em sala de aula.