Experimental biochemistry
- UE code MPHAB203
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Schedule
25Quarter 2
- ECTS Credits 3
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Language
French
On completion of the course, the student will be able to:
• Use appropriately laboratory equipment and materials used in biochemistry (balance, pH meter, spectrophotometer, centrifuge, pipette, micropipette, spectrophotometric cuvette, 96-well plate, pH paper, Pasteur pipette, glassware, separation column, filter).
• Prepare a solution of a defined concentration from a powder or liquid.
• Carry out a dilution.
• Prepare a buffer and understand its purpose.
• Perform a calibration curve and a spectrophotometric assay.
• Quantify the activity of an enzyme by spectrophotometric quantification of a product or substrate.
• Purify a protein of interest by ion exchange chromatography and differential centrifugation.
• Write a report on the experience.
• Analyse raw experimental data and present it using a spreadsheet (Excel).
• Interpret and critically evaluate experimental data.
• Develop the ability to manage time and work in pairs.
• Restate the name, 1-letter code, 3-letter code, structure, pKa and class (according to the polarity of the side chain) of the amino acids.
• Restate the characteristics of the peptide bond and know how to draw a peptide with a correct ionisation according to a defined pH.
• Restore the structural levels of proteins.
• Understand and reproduce the theoretical principles of the following separation techniques: ion exchange chromatography, electrophoresis and molecular exclusion chromatography.
• Draw the 3 linearizations used in enzymology and to know how to calculate the enzymological kinetic constants from these linearizations.
• Identify the type of inhibition of an enzyme inhibitor from its impact on kinetic constants.
• Develop an enzymological assay protocol by choosing the appropriate equipment and method for its implementation.
This course will cover:
• Preparation of solutions and spectrophotometric exercises.
• Structural characteristics of amino acids and proteins.
• Chemical characteristics and separation techniques for amino acids and proteins.
• Enzymology exercises.
• Development of an enzymology protocol: study of the enzymatic kinetics of alkaline phosphatase.
The assessment is carried out during the four-month period as well as by an examination in the examination session through 3 marks:
• Grading of the practical reports. This mark accounts for 10% of the final mark.
• Laboratory behaviour and practical examination. This mark accounts for 40% of the final mark.
• Written examination in session on the material covered during the practical and tutorial sessions. This mark accounts for 50% of the final mark.
Any failure (grade <10) in 2 of these grades will result in a direct failure of the entire subject of that unit. Attendance at practical sessions is compulsory. Any unjustified absence does not allow the written examination to be taken.
Training | Study programme | Block | Credits | Mandatory |
---|---|---|---|---|
Bachelier en sciences pharmaceutiques | Standard | 0 | 3 | |
Bachelier en sciences biomédicales | Standard | 0 | 3 | |
Bachelier en sciences pharmaceutiques | Standard | 2 | 3 |