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.

Pre-requisites

Organic Chemistry [MCHIB107]

Co-requisites

General biochemistry [MPHAB202]

Teaching methods

The student will have at his disposal:

• A syllabus of tutorials.

• A syllabus of practical work.

• The powerpoint materials used in the tutorials.

• Videos illustrating theoretical and/or practical points.

The aim of the tutorial sessions is to provide the theoretical and practical material necessary for the proper conduct and understanding of the practical sessions and the processing of the results generated during these sessions.

Assessment method

The course consists of two components: a practical component and a theoretical component.

Practical Component

The practical component includes an introductory session and an initial lab (Lab 1), which are not graded, as well as three graded labs (Lab 2, Lab 3, and Lab 4). Grading is based on pre-lab quizzes, post-lab reports, and a practical exam administered outside of exam periods.

The grade for the practical component is broken down as follows:

Lab 2: 15%;

Lab 3: 15%;

Lab 4: 15%;

practical exam held outside the regular exam period: 55%.

Attendance at lab sessions is mandatory. If a student misses more than one session, they will not be permitted to take the practical exam outside the exam period. This exam is administered only during the first exam period and cannot be retaken during the third exam period.

Theoretical Component

The theoretical component consists of tutorials (TD) and a theoretical exam administered during the exam session. This exam constitutes the entire grade for the theoretical component.

Assessment Procedures for the First Exam Session

In the first exam session, the final grade for the course unit corresponds to the weighted average of the two components:

practical component: 40%;

theoretical component: 60%.

To pass the course unit, students must earn a grade of at least 10/20 in each of the two components. If the grade earned in either component is less than 10/20, the final grade for the course unit is capped at 8/20, regardless of the weighted average obtained.

Assessment Procedures for the Third Session

In the third session, only the theoretical exam may be retaken. The grade earned on this exam then constitutes the entire final grade for the course unit. The grade for the practical component and the weighting applied in the first session are therefore no longer taken into account.

Language of instruction

French