DSP (down 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".

DSP or DownStream Processing are the downstream processes in the biomedicine production chain. They correspond to the extraction and purification of the molecule of interest (recombinant protein or monoclonal antibody derived from a hybridoma). Distribution/packaging and related controls will also be covered.

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 have theoretical and practical knowledge of the operation of cell lysis, chromatography and distribution/conditioning equipment. They will be able to adapt the choice of equipment and processes to the characteristics of the target protein.

Initially, students will study the physico-chemical properties of the target protein using simple sequence analysis tools. Based on these data and the final use of the protein, a purification strategy will be proposed by the students (TD).

These elements will be put into practice in the practical part of the course, in which students will purify the target protein while carrying out a series of sampling and on-line process controls to ensure traceability and final product quality.

Secondly, the students will have 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 to therapeutic proteins. They will have to determine production yields, degree of purity, nature of impurities and confirm the identity of the target protein (TD).

This UE is based on 2 cross-disciplinary projects on skills blocks 1(Mobilizing the 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). Purification, assay and controls of the antibody will be carried out following cell culture (UE2.2) and controls of the producing clone (UE 1.3).

Project C: Production and purification of a fusion protein used as a veterinary vaccine in prokaryotic hosts in fed-batch bioreactors (pilot scale). Following the USP practical part (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 storage conditions for the protein produced (stability, sterility, etc.). All results, analyzed critically, can be presented in the form of a batch production report.

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

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

- Principles of protein purification

- Prepare and run a biotechnological purification process

- Characterizing a therapeutic protein for industrial biomanufacturing

- Prepare and discuss a batch report