USP (upstream process)

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

USP or upstream processing refers to the early stages of the production chain for a biopharmaceutical product. It involves the production of cells (cell therapy) and products (proteins, vaccines, monoclonal antibodies, etc.) in bioreactors.

The program covers:

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

2/ Management of bioproduction processes.

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

After presenting a standard bioreactor (mechanically agitated and aerated tank), the different scales and the concept of scale-up will be discussed, as well as other types of bioreactors (bubble column, airlift, etc.).

Next, all the preparatory steps prior to culture will be described and carried out both in the form of tTD and during practical work on the Bioproduction teaching platform. This part will cover: the preparation of culture media and the adaptation of their composition, assembly, verification, calibration of probes and the bioreactor, sterilization, implementation of an aseptic sampling system, and preparation of the inoculum (cell banks will be covered in EU 1.3 - Recombinant protein expression systems).

Next, the focus will be on "How to conduct and implement a culture in a bioreactor."

In this section, we will discuss:

- Indicators for monitoring biological reactions and monitoring (offline, near-line, and online techniques) and crop management. The program includes: control loops, an introduction to PAT (Process Analytical Technology) and its integration with quality procedures (e.g., implementation of monitoring indicators, control charts, etc.).

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

- Different operating modes (batch, fed-batch, continuous): after a general presentation of the different modes, each one is examined in detail. The aim is for students to become involved in an engineering approach to developing viable technical solutions so that they are able to:

o Analyze experimental results

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

o To anticipate the various possible scenarios for feed management for fed-batch and continuous cultures

o To carry out crops during practical work on the educational platform in Bioproduction

This EU is based on three projects that cut across skill blocks 1 (Mobilizing key theories and concepts for bioproduction), 2 (Preparing, designing, and conducting a bioproduction process), and 3 (Communicating about one's work), namely:

Project A: Production and purification of a monoclonal antibody in hybridoma (laboratory scale – screening). Cell culture (preparation of media, cell counting and culture) will be carried out after the producer clone has been checked (UE 1.3).

Project B: Production of a fluorescent recombinant fusion protein in a prokaryotic host in batch mode bioreactors (laboratory scale). Cultivation and monitoring will be carried out after the construction of the producer strains and their characterization on a small scale (UE 1.2). In addition to carrying out the cultivation and analyzing the results obtained, this practical work aims to master the preparation of a bioreactor (calibration, sterilization, etc.). This practical work is also linked to UE 2.1 for the establishment of the experimental plan and to UE 3.1 for the production of reporting documents.

Project C: Production and purification of a fusion protein used as a veterinary vaccine in a prokaryotic host in fed-batch mode bioreactors (pilot scale). Preparation of the fermenter and fed-batch culture from a cell bank of anE. coli strain. Monitoring of the culture (online and offline parameters) and recovery of biomass, calculation of yields. The complete DSP will be carried out in EU 2.3. And batch records/production reports will be produced in conjunction with EU 2.1 and 3.1.

The educational platform for bioproduction:

USP:

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

- 4 instrumented benchtop bioreactors (2 to 5 L capacity)

- 1 pilot-scale instrumented bioreactor (20 L usable capacity)

DSP:

- Centrifuge

- High-pressure homogenizer (Emulsiflex, Avestin)

- AKTAPure + preparative GSTPrep affinity column

- Instrumented pilot freeze dryer

See the EU description

- Prepare a bioreactor

- Cultivating and implementing a culture in a bioreactor

For more details, refer to the EU description.