EC
USP Development practicals
Description
Objectives in terms of knowledge (course content) :
General principles of bioproduction and purification of recombinant proteins in E. coli.
Search for optimum expression conditions of different proteins in E. coli using different induction methodologies such as auto-induction and induction with IPTG at different temperatures (37°, 30° or 20°).
Scale-up of the retained protein and purification of the protein by chromatography, such as Maltose Binding Protein (MBP), Chelate (His-Tag) or Gluthation-S-Transferase. (GST), and characterization of the purified protein; estimation of purity, quantification, yield obtain etc.
Compétences visées
Objectives in terms of skills :
Reflection and criticism on recombinant protein production in E. coli, scale-up and adapted methods for their purification.
Reflection on the experiments and controls to be carried out to validate the project
Ability to use different chromatography techniques depending on the components to be purified.
Project management and critical analysis of experimental results
Team Work Management
Bibliographie
Lecture PDF, Publications, protocoles, G healthcare documentations on chromatography technics. Technical sheets of the products used.