DSP (downstream 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.”

DSP (Downstream Processing) processes are the so-called downstream processes in the production chain of a biopharmaceutical. They involve the extraction and purification of the target molecule (recombinant protein or monoclonal antibody derived from a hybridoma). In this course unit, the aspects of distribution and packaging, as well as the related controls, will also be covered.

The program focuses on:

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

2) Process management and quality control for the production of recombinant proteins

Students will gain theoretical and practical knowledge of how cell lysis, chromatography, and fractionation/packaging equipment works. They will be able to select the appropriate equipment and processes based on 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 protein’s intended use, the students will propose a purification strategy (lab).

These elements will be applied in the practical portion of the course, during which students will purify the target protein while performing a series of in-process sampling and quality control checks to ensure traceability and the final quality of the product.

Next, students will need to evaluate the performance of the process and the quality of both the active ingredient and the finished product by proposing an analytical approach appropriate for therapeutic proteins. They will thus need to determine production yields, purity levels, and the nature of impurities, and confirm the identity of the target protein (TD).

This learning unit is based on two cross-cutting projects covering skill sets 1 (Applying Key Theories and Concepts to 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 using hybridoma cells (laboratory scale—screening). Purification, titration, and testing of the antibody will be performed 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 USP practical session (UE.2.2), the resulting biomass is weighed and then lysed (HHP). Emphasis is placed on monitoring critical parameters (viscosity, pressure, concentration, etc.), the efficiency of the chosen process (yield, purity), and the storage conditions for the produced protein (stability, sterility, etc.). All results, analyzed critically, may be presented in the form of a batch production report.

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

- 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