USP (up stream process)

This teaching unit (UE) is part of block 2 of the skills targeted by the L3 Bio²S, i.e."Preparing, designing and running a bioproduction process".

USP or Upstream processing are the "upstream" processes in the biomedicine production chain. They correspond to the production of cells (cell therapy) and products (proteins, vaccines, monoclonal antibodies, etc.) in bioreactors.

The program covers :

1/ Knowledge of equipment: bioreactors from laboratory to pilot or industrial scale, traditional and single-use systems and their direct environment (sensors, etc.).

2/ Managing bioproduction processes.

Students will have a theoretical and practical knowledge of the structure and function of a bioreactor and how to prepare it for cultivation. structure and function of a bioreactor and how to prepare it for cultivation..

After a presentation of a standard bioreactor (mechanically stirred and aerated tank), the different scales and the notion of scale-up will be discussed, as well as other types of bioreactor (bubble column, Airlift, etc.).

All the preparatory stages prior to culture will then be described and carried out, both in the form of tutorials and practical work on the Bioproduction teaching platform. This part will cover: preparation of culture media and adaptation of their composition, assembly, checking, calibration of probes and bioreactor, sterilization, setting up an aseptic sampling system, inoculum production (cell banks will be covered in UE 1.3- Recombinant protein expression systems).

The focus will then be on "How to run and implement a bioreactor culture".

In this section, we will develop :

- Biological reaction monitoring indicators and monitoring (off-line, near-line, on-line techniques) and crop management. The program includes: control loops, an introduction to the PAT (Process Analytical Technology) approach and its coupling with the quality approach (e.g.: setting up monitoring indicators, control charts, etc.).

- Management of industrial production constraints (in particular limitations due to transfer phenomena such as heat and gas (_O2))

- The different operating modes (batch, fed-batch, continuous): after a general presentation of the different modes, a focus on each of them is proposed. The aim is for students to integrate an engineering approach to the development of viable technical solutions, and to be able to :

o Analyze experimental results

o Use a production planning tool (simulation) on an industrial scale (or experimentation on a pilot lab scale).

o Forecast the various possible feed management scenarios for fed-batch and continuous crops.

o Produce cultures during practical work on the Bioproduction teaching platform

This UE is based on 3 cross-disciplinary projects related to skills blocks 1(Mobilizing key theories and concepts for bioproduction), 2(Preparing, designing and running a bioproduction process) and 3(Communicating about one's work), i.e. :

Project A: Production and purification of a monoclonal antibody in hybridoma (laboratory scale -screening). Cell culture (preparation of media, counting and culturing of cells) will be carried out after testing of the producing clone (EU 1.3).

Project B: Production of a recombinant fluorescent fusion protein in prokaryotic hosts in bioreactors in batch mode (laboratory scale). Cultivation and monitoring will be carried out after the production strains have been constructed and characterized on a small scale (UE 1.2). In addition to carrying out the culture and analyzing the results obtained, the aim of this practical course is to master the preparation of a bioreactor (assembly, calibration, sterilization, etc.). This lab is also linked to UE 2.1 for experimental design, and to UE 3.1 for reporting documents.

Project C: Production and purification of a fusion protein used as a veterinary vaccine in prokaryotic hosts in fed-batch bioreactors (pilot scale). Preparation of the fermenter and fed-batch culture of anE. coli strain from a cell bank. Culture monitoring (on-line and off-line parameters) and biomass recovery, yield calculation. The complete PSD will be carried out in UE 2.3, while batch records/production reports will be produced in conjunction with UE 2.1 and 3.1.

The bioproduction teaching platform :

USP :

- Eukaryotic and prokaryotic cell culture (type II SHP, incubators)

- 4 instrumented benchtop bioreactors (2 to 5 L)

- 1 pilot-scale instrumented bioreactor (useful 20 L)

DSP :

- Centrifuge

- High-pressure homogenizer (Emulsiflex, Avestin)

- AKTAPure + GSTPrep preparative affinity column

- Instrumented pilot freeze-dryer

See the EU description

- Preparing a bioreactor

- Manage and implement bioreactor culture

For further details, please refer to the EU description