Learning outcomes

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.

Content

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.

Assessment method

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.

 

Language of instruction

French