2023-2024 / BIOC0735-1



60h Th, 45h Pr, 15h AUTR

Number of credits

 Master in bio-informatics and modelling (120 ECTS)7 crédits 
 Master in biochemistry and molecular and cell biology (120 ECTS)7 crédits 


Alain Brans, Patrick Fickers, Moreno Galleni, Sylvie Legrand, André Matagne, N..., Claire Remacle, Mohammed Terrak, Pierre Tocquin, Marylène Vandevenne


Moreno Galleni

Language(s) of instruction

French language

Organisation and examination

Teaching in the second semester


Schedule online

Units courses prerequisite and corequisite

Prerequisite or corequisite units are presented within each program

Learning unit contents

Teaching unit contents

This module will explain the principles of the methods used to produce and purify biological macromolecules and to characterize their biochemical properties. Exercises and practical work will be organized to illustrate recent methods for producing biomolecules in different hosts, techniques for their optimal expression in fermenters, and techniques for purifying and characterizing the catalytic properties of enzymes.

This module consists of four parts:


i) Recombinant protein expression systems

The aim of this section is to present the various heterologous protein expression systems.

1) Recombinant protein production (Prokaryotes)

2) Recombinant protein production (yeast and filamentous fungi)

3) Recombinant protein production (plants)

4) Recombinant protein production in microalgae

5) Recombinant protein production (mammalian cell and Baculovirus)?


Design of an in Silico production plasmid

Recombinant bacterial strain

Production of recombinant proteins in plant systems (8h TP)

ii) Bioreactor and Down processing

1) Bioreactor culture

2) Downstream processing


Development of a microbial process

iii) Production and purification of proteins and metabolites

1) Review of chromatography theory

2) Sample preparation

3) Exclusion chromatography

4) Ion exchange chromatography

5) Hydrophobic chromatography

6) High-performance liquid chromatography (HPLC)

7) Affinity chromatography

8) Electrophoresis and capillary electrophoresis

9) Analyte separation using non-chromatographic techniques

Practical work

Purification of pike paravalbumin

Day 1: Sample preparation

           Heat fractionation and ammonium sulfate

           Sample dialysis

Day 2: Preparation of molecular sieve columns and ion exchanger

           Sample deposition on molecular sieve

Day 3: Identification of paravalbumin 1

           Sample concentration

Day 4: Sample deposition on cation exchanger

           Isolation of paravalbumin II and III

Day 5: UV/Vis spectra of paravalbumin samples

          Denaturing polyacrylamide gel


A visit to a biotechnology company will be organized.

iv) Enzymology

Chapter 1: General properties of enzymes.

Chapter 2: Enzymatic kinetics. Stationary state - Independent sites.

Chapter 3: Inhibition.

Chapter 4: Influence of physico-chemical parameters

Chapter 5: Two-state systems.

Chapter 6: Transient states.

Chapter 7: Cooperative systems.

Chapter 8: Inactivators.

Chapter 9: Cofactors.

Chapter 10: The mechanism of action of chymotrypsin.

Chapter 11: Catalysis: hypotheses of the mechanism of action.

Chapter 12: Control of enzymatic activity.

Chapters 1 to 3 are covered in Bac 3. Chapters 8 and 9 are part of the course notes but are not detailed in the oral course. They are given for information purposes and can be considered as documentation. Chapters 7 and 12 are given in the form of tutorials.

In addition, one session will be reserved for a lesson on the formation of disulfide bridges in proteins, given by Professor Jean-François Collet. The aim is to illustrate the wide variety of biochemical reactions catalyzed by enzymes.

Practical work and tutorials

An exercise session will be held in connection with chapter 6.

Chapters 7 and 12 are given as tutorials.


Visits by small groups (2 to 4 students) to the Robotein technology platform. The "Dynamic Light Diffusion" technique will be presented.


Learning outcomes of the learning unit

At the end of the theoretical courses given in the "Biotechnology" module, students will be able to :

  • master the different molecular systems for recombinant protein production
  • master the basic principles of bioreactor culture and downstream processing
  • develop a protocol for purifying proteins and metabolites
  • Characterize the kinetic properties of an enzymatic reaction
  • Determine the most appropriate methodological approaches according to the nature of the biomolecules to be characterized and their uses
At the end of the practical/directed work, students will be able to:

  • Use data generated by a fermenter during protein expression by a recombinant organism.
  • Define the genetic elements involved in the design of plasmids for heterologous protein production.
  • Understand and explain the behavior of biomolecules on chromatographic supports.
  • Use IT tools to monitor cell behavior in the fermenter and the level of heterologous protein production.
  • Analyze and interpret a set of kinetic data.

Prerequisite knowledge and skills

This module will build on the concepts covered in the following courses:

Bacteriology (MICR072-1), Molecular Biology (GENE9002 and GENE9003), Chemistry of Biological Macrolécules (BIOC09242), Equilibrium in Biochemistry and Enzymatic Kinetics (BIOC9243) and the "Nucleic Acids", "Proteins" and "Structural Biology" toolboxes.

Planned learning activities and teaching methods

Theoretical lessons will last two hours. They will mainly consist of ex-cathedra lectures with active participation by students, through the resolution of small exercises relating to the essential concepts (specified during the lessons) covered in previous lectures. Attendance is recommended, and note-taking is encouraged.

For the "Recombinant Protein Expression System" part of the course, students will have the option of working on computer-based practical sessions. For the other sections, students will form groups of 2 to 4 and carry out a series of experiments designed to put into practice the theoretical concepts covered in the course.

Mode of delivery (face to face, distance learning, hybrid learning)

Face-to-face course

Recommended or required readings

Powerpoint presentations, TPs/TDs notes and exercises from theory classes will be uploaded to e-Campus.

Exam(s) in session

Any session

- In-person

written exam ( open-ended questions )

Additional information:

This module will be assessed by means of a written examination covering the various parts of the course. It will assess the knowledge acquired, as well as the ability to develop a cross-disciplinary analysis.

The written exam will include open-ended questions on protein production, purification and characterization. They will cover the following topics: heterologous expression systems, purification methods and downstream processing. Finally, open questions and/or exercises from the "Enzymology" section will be proposed.

The grade for this module will correspond to the arithmetic sum of the 4 partims.

Work placement(s)

Organisational remarks and main changes to the course

Practical work and/or tutorials are compulsory. The concepts covered are likely to form part of the assessment. All absences must be justified, and students are required to put their work in order. If a report is required, it must be completed even in the event of absence.


The supervisors are available to answer your questions: either during the lectures/practical sessions given as part of this thematic module, or by e-mail (during office hours and no later than 2 days before the exam or report submission).


Module coordinator :

Galleni Moreno

Tel : 04/366.35.49 
e-mail : mgalleni@uliege.be


Co-tutors :

Dr. Brans Alain (abrans@uliege.be)
Dr. Terrak Mohamed (mterrak@uliege.be)
Prof. Fickers Patrick (pfickers@uliege.be
Dr. Legrand Sylvie (S.legrand@uliege.be)
Prof. Remacle Claire (Claire.Remacle@uliege.be)
Dr. Tocquin Pierre (ptocquin@uliege.be)
Prof. Matagne André (amatagne@uliege.be)
Dr. Vandevenne Marylène (mvandevenne@uliege.be)

Association of one or more MOOCs