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An extensive analysis of a protein is necessary to establish a well characterized biologic, independent of whether the protein is later used for research, as pre-clinical material for animal experiments, or whether the protein will be the active ingredient of a marketed product. Protein characterization always has to include physicochemical, biochemical and cell-based analyses.
Protein analytics is also a pivotal part of downstream process development. In-process control, the demonstration of DNA removal and host cell impurities and the final analysis of the protein has to be carried out at the highest quality level.
Scil Proteins Services employs leading-edge technologies and instrumentation that is complemented by skilled project managers who routinely perform in-process control (IPC) and analysis of final products.
Methods of protein characterization at Scil Proteins comprises: |
Methods of physicochemical protein analysis for the determination of the degree of purity, concentration and structural properties at Scil Proteins Services include:
- Electrophoresis (SDS-PAGE, Western Blot)
- HPLC (reversed phase, size exclusion, ion exchange)
- Analytical ultracentrifugation
- Spectroscopy (UV-VIS, fluorescence, circular dicroism)
- N-terminal sequencing
- Mass spectrometry
Further approches of protein characterization comprises:
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Methods for biochemical analysis at Scil Proteins Services include:
- Determination of DNA contamination and host cell impurities (Threshold™ system)
- Determination of the endotoxin content
- Biacore analysis
- Enzyme assays to determine and quantify the catalytic activity of the target protein
- Determination of the protein content and activity by ELISA
- Immunohistochemical staining
- Immunofluorescence cell labelling
Further approches of protein characterization comprises:
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Scil Proteins Services has developed and established a broad variety of assays to determine the biological activity of the target proteins in living cells.
Examples are:
- WST assay for vitality and cytotoxicity
- Brom deoxyuridine (BRDU) assay for proliferation
- Boyden-chamber-assay to determine the cell migration and invasion behavior as a response on attractants
- PC-12 assay for neurogenic differentiation
- Osteochondrogenic differentiation assay using mesenchymal stem cells
- Determination of aminoglucans and isolation of RNA by 3-D culturing of human primary chondrocytes
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