Upstream Process Development
Upstream process development at Glycotope offers biotechnological contract research and development based on mammalian cell culture systems with a special expertise on perfusion cultivation technologies.
We offer ISO 9001 compliant services to ensure controlled quality of biopharmaceutical proteins.
Beside development in customers’ own cell lines, we offer the GlycoExpress® (GEX®) technology, a well-established human expression platform for the screening and production of biotherapeutics:
- Screening of glycosylation variants produced in different GEX® cells
- Cell line development in customers’ own cell line or in GEX® cells
- Clone screening and process optimization in microscale bioreactors in batch, fed-batch or perfusion mode.
- Upstream process development at 1-10 L scale for batch, fed-batch or perfusion (Xcell™ ATF or Centritech®) operation.
- Track record of successful process development for mAbs (including defucosylatad Abs), antibody constructs, hormones, blood factors and other glycooptimized or difficult-to-express proteins.
- High yield expression combined with high reproducibility and scalability.
Process Development Services
Our process development services are enhanced by extensive work in immunology, cell biology, molecular biology and protein chemistry. From intense work on the development of our own pipeline products we have acquired a very broad portfolio of analytical methods. This we bring in into our service offering for our clients.
Process Transfer and Process Development
We develop upstream processes (USP) and downstream processes (DSP) using our platform or a customized variation. Processes can also be transferred from any other development or manufacturing site for improvement. We are experts in perfusion and fed-batch processes, including their establishment and characterization.
We use all state-of-the-art technologies (2 Ambr-systems, batteries of small-scale bioreactors from 1 to 10 L working volume) in USP development. Machines from GE (Äkta Avant and Pure series) are available in our laboratories for protein purification development.
Development of purification processes
In the development of downstream processes (DSP) the structural and functional parameters of the recombinant protein must be retained while simultaneously all potential impurities are removed.
We perform method development for protein purification processes including filtration (cell separation and TFF/diafiltration development) and capturing from harvest, through scouting for convenient chromatography methods (IEC, SEC, HIC, RP, multimodal and affinity) in order to achieve an efficient removal of host cell proteins (HCP), culture medium components, endotoxins and host cell DNA as critical impurities.
Denaturation of proteins during long-term storage must be avoided. Most prominent problems are aggregation, fragmentation, deamidation, and oxidation. Glycotope uses accelerated stress studies for the liquid or lyophilized storage buffer to optimize the stability of your protein.
Our core Process Development competencies
- Process development ready for tech-transfer and scale-up to pilot and production level
- Optimization of existing processes
- DoE, robustness tests
- Formulation development
- Stability studies under R&D
- Considering purity demands in accordance with GMP guidelines
Analytical Services and Stability Studies
Glycotope has proven experience in customized bio-analytical method development, validation and routine application.
Glycotope’s analytical services strongly support customers for specific biological assay development at any point along the product development path. We provide ISO9001-compliant analytical services and work together with our clients to develop and set-up efficient first-class quality control management to ensure the highest product quality and to help reduce the overall time and cost of product development.
Stability studies & formulation development
For demonstration and documentation of the batch-to-batch consistency of any given biopharmaceutical material, its identity, purity, quantity, bioactivity and profile of impurities must be confirmed. These tasks are addressed by our portfolio of analytical routine methods for quality control of a recombinant protein. Glycotope GmbH provides comprehensive expertise in the development and validation of analytical assays.
Stability data can be generated under ISO 9001 according to the ICH guidelines. Accelerated stability data can be used for formulation development.
The services include:
- Development of various analytical services including identity, purity and potency testing
- cGMP compliant method validation and transfer
- Performance of stability studies according to ICH guidelines
- HPLC (SEC, IEC, RP)
- Endotoxin content (LAL-test, Haemotox rFc, EndoLISA®)
- Host cell protein (generic kits, GEX®-specific ELISA, or development of customized tests)
- Process related impurities, e.g. Protein A, HSA, Insulin (ELISA)
- Bradford, BCA
- UV/Vis spectroscopy
- HPLC (SEC) and FPLC (Affinity, e.g. analytical Protein A measurement)
- Western blotting
- Isoelectric focusing, WCX
- Mass spectrometry
- Peptide Mapping
- Mono- and Oligosaccharide analysis
- Bioactivity assays
- Antigen ELISA
- Binding kinetics using DRX (Dynamic biosensors; substitution of Biacore SPR technology)
- Other specific binding assays: For more information please see our page “bioactivity and cellular assays”
- Ion chromatography