USP (upstream process)

This course unit (UE) is part of Skill Block 2 of the Bio²S Bachelor’s program (L3), titled “Preparing, Designing, and Conducting a Bioproduction Process.”

USP (Upstream Processing) refers to the “upstream” processes in the production chain of a biopharmaceutical. These processes involve the production of cells (cell therapy) and products (proteins, vaccines, monoclonal antibodies, etc.) in bioreactors.

The program focuses on:

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

2/ Management of bioproduction processes.

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

Following a presentation on a standard bioreactor (a mechanically agitated and aerated tank), the various scales and the concept of scale-up will be discussed, along with other types of bioreactors (bubble column, airlift, etc.).

Next, all the preparatory steps prior to culture setup will be described and carried out both in the form of tTDs and during hands-on lab sessions on the Bioproduction educational platform. This section will cover: the preparation of culture media and the adjustment of their composition; the assembly, verification, and calibration of the probes and the bioreactor; sterilization; the setup of an aseptic sampling system; and the preparation of the inoculum (cell banks will be covered in UE 1.3—Recombinant Protein Expression Systems).

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

In this section, the following topics will be covered:

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

- Management of industrial production constraints (particularly limitations caused by transport phenomena such as heat and gas (_O2))

- The different operating modes (batch, fed-batch, continuous): Following a general overview of the different modes, the course provides an in-depth look at each one. The goal is for students to engage in an engineering approach to developing viable technical solutions so that they will be able to:

o Analyze experimental results

o Use a production planning (simulation) tool at the industrial scale (or conduct experiments at the lab-pilot scale)

o To anticipate the various possible scenarios for feed management in fed-batch and continuous-flow processes

o To perform cultures during lab sessions on the Bioproduction educational platform

This learning unit is based on three cross-cutting projects spanning competency blocks 1 (Applying Key Theories and Concepts to Bioproduction), 2 (Preparing, Designing, and Implementing a Bioproduction Process), and 3 (Communicating About One’s Work), namely:

Project A: Production and purification of a monoclonal antibody using hybridoma cells (laboratory scale—screening). Cell culture (preparation of media, cell counting, and cell culture) will be performed after the producer clone has been verified (Work Package 1.3).

Project B: Production of a fluorescent recombinant fusion protein in a prokaryotic host using batch-mode bioreactors (laboratory scale). Culture setup and monitoring will be performed after the construction of the producer strains and their small-scale characterization (Module 1.2). In addition to conducting the culture and analyzing the results obtained, the objective of this lab is to master the preparation of a bioreactor (assembly, calibration, sterilization, etc.). This lab is also linked to Unit 2.1 for the design of the experimental plan and to Unit 3.1 for the preparation of reporting documents.

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

The educational module on bioproduction:

USP:

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

- 4 instrumented benchtop bioreactors (2 to 5 L usable volume)

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

DSP:

- Juicer

- High-pressure homogenizer (Emulsiflex, Avestin)

- AKTAPure + GSTPrep preparative affinity column

- Instrumented pilot-scale freeze-dryer

See the EU description

- Set up a bioreactor

- Cultivate and grow a culture in a bioreactor

For more details, see the course description