DSP (downstream 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."

DSP or DownStream Processing procedures are the downstream procedures in the production chain of a biopharmaceutical. They involve the extraction and purification of the molecule of interest (recombinant protein or monoclonal antibody derived from a hybridoma). This EU will also cover distribution/packaging aspects and related controls.

The program covers:

1) Knowledge of equipment (cell lysis, chromatography, filtration, freeze-drying, etc.)

2) Process management and quality control of recombinant proteins produced

Students will acquire theoretical and practical knowledge of how cell lysis, chromatography, and distribution/packaging equipment works. They will be able to adapt the choice of equipment and processes according to the characteristics of the target protein.

First, students will study the physicochemical properties of the target protein using simple sequence analysis tools. Based on this data and the end use of the protein, students will propose a purification strategy (tutorial).

These elements will be implemented in the practical part of the course, in which students will purify the target protein while performing a series of online samples and checks on the process to ensure traceability and the final quality of the product.

In a second step, students will be required to evaluate the performance of the process and the quality of the active ingredient as well as the finished product by proposing an analytical approach appropriate for therapeutic proteins. They will thus be required to determine production yields, purity levels, the nature of impurities, and confirm the identity of the target protein (TD).

This EU is based on two cross-disciplinary projects covering 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). Purification, titration, and testing of the antibody will be carried out following cell culture (UE2.2) and testing of the producing clone (UE 1.3).

Project C: Production and purification of a fusion protein used as a veterinary vaccine in a prokaryotic host in fed-batch bioreactors (pilot scale). Following the practical part of USP (UE.2.2), the biomass obtained is weighed and then lysed (HHP). Emphasis is placed on controlling critical parameters (viscosity, pressure, concentration, etc.), the efficiency of the chosen process (yield, purity), and the storage conditions of the protein produced (stability, sterility, etc.). All results, analyzed critically, may be presented in the form of a batch production report.

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

- SDS-PAGE and Western blot, UV spectroscopy, nanoDSF

- Understand the principles of protein purification

- Prepare and conduct a biotechnological purification process

- Characterize a therapeutic protein in the context of industrial bioproduction

- Prepare and discuss a batch report