Development of production methods for recombinant protein
- recombinant proteins produced in E.coli (cytoplasmic or periplasmic expression) and P. pastoris (secretion).
- Construction of protein expression systems (E. coli or P. pastoris strains):
- vectors design including selection between different strains, vectors, selection markers, promoters, secretion signals, coding sequences, modes of codon optimization.
- transformation, clones screening and selection of the best producing clones.
- screening of optimal process conditions
- Harvest and extraction:
- optimization of the harvest conditions for cells or media recovery
- development of the extraction conditions: high pressure treatment, osmotic shock, chemical lysis
- preparation, solubilization and refolding of inclusion bodies
- high throughput screening of resins and chromatographic conditions
- validation of resins and fine-tuning of process conditions using small-scale columns
- Ultrafiltration and microfiltration:
- membranes and process conditions screening
- Industrial transfer:
- scaling-up studies (chemical engineering)
- multi-parameters process monitoring
- Research Cell Banks (50 vials) with certificate of analysis
- Purified material for in vitro trials (activity testing, formulation studies…).
- Development report including:
- method of production
- operating parameters at the different scales of production
- A process transferable to the Xpress Biologics GLP platform.
- A process “ready-to-transfer” to GMP manufacturing.