An optical biosensor for monitoring recombinant proteins in process media
| Autor: | Darrin M Disley, Christopher R. Lowe, Sproule Kenneth, Paul R. Morrill |
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| Rok vydání: | 1999 |
| Předmět: |
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Molecular Optics and Photonics Biomedical Engineering Biophysics Analytical chemistry engineering.material Ligands Protein Structure Secondary chemistry.chemical_compound Coating Monolayer Electrochemistry Humans Insulin Sulfhydryl Compounds Surface plasmon resonance Mercaptoethanol Chromatography Chemistry Surface plasmon Self-assembled monolayer General Medicine Surface Plasmon Resonance Ligand (biochemistry) Recombinant Proteins Dextran engineering Gold Biosensor Biotechnology |
| Zdroj: | Biosensors and Bioelectronics. 14:481-493 |
| ISSN: | 0956-5663 |
| DOI: | 10.1016/s0956-5663(99)00021-4 |
| Popis: | This paper describes the construction of a sensor for the direct monitoring of a recombinant protein, the human insulin analogue (MI3). The surface plasmon resonance (SPR) sensor incorporates an immobilised, sterilisable affinity-ligand that has been designed to bind to MI3. In practice, gold SPR devices were fabricated with; a 2D assembly of ethanethiol-modified ligand, a 2D mixed-assembly of ethanethiol-modified ligand and mercaptoethanol, a 3D coating of ligand-modified terminal-thiolated poly(vinyl)alcohol (PVA) or a 3D hydrogel of dextran coupled to a self-assembled monolayer (SAM) of mercaptohexaneundecanl-ol. Routine measurement of the concentration MI3 in the concentration range 1-100 mg/l in pilot-scale samples of crude fermentation broth have been achieved with high sensitivity levels and a high signal-to-noise ratio. Analysis can be achieved within10 min with the active surface being regenerable for at least 60 cycles over a 6 month period. The coupling of a robust, sterilisable and highly-selective sensor-coating with suitable transducer technologies promises to deliver sensors that are capable of direct in situ monitoring of biopharmaceuticals in industrial bioprocesses. |
| Databáze: | OpenAIRE |
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