Utilization of newly developed immobilized enzyme reactors for preparation and study of immunoglobulin G fragments
| Autor: | Michal Holèapek, Jiøı́ Lenfeld, Josef Královský, Lucie Korecká, Roxana Cecal, Milan J. Beneš, Jean-Louis Viovy, Michael Przybylski, Zuzana Bílková, Nicolas Minc |
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| Rok vydání: | 2004 |
| Předmět: |
Immobilized enzyme
Clinical Biochemistry Molecular Sequence Data Biochemistry Immunoglobulin G Mass Spectrometry Analytical Chemistry chemistry.chemical_compound Bioreactors Spectroscopy Fourier Transform Infrared medicine Amino Acid Sequence Immunoglobulin Fragments Chromatography High Pressure Liquid chemistry.chemical_classification Chymotrypsin Chromatography biology Chemistry Proteolytic enzymes Cell Biology General Medicine Trypsin Enzymes Papain Enzyme biology.protein Electrophoresis Polyacrylamide Gel Digestion medicine.drug |
| Zdroj: | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences. 808(1) |
| ISSN: | 1570-0232 |
| Popis: | The newly developed immobilized enzyme reactors (IMERs) with proteolytic enzymes chymotrypsin, trypsin or papain were used for specific fragmentation of high molecular-mass and heterogeneous glycoproteins immunoglobulin G (IgG) and crystallizable fragment of IgG (Fc). The efficiency of splitting or digestion were controlled by RP-HPLC. The specificity of digestion by trypsin reactor was controlled by MS. IMERs (trypsin immobilized on magnetic microparticles focused in a channel of magnetically active microfluidic device) was used for digestion of the whole IgG molecule. The sufficient conditions for IgG digestion in microfluidic device (flow rate, ratio S:E, pH, temperature) were optimized. It was confirmed that the combination of IMERs with microfluidic device enables efficient digestion of highly heterogeneous glycoproteins such as IgG in extremely short time and minimal reaction volume. |
| Databáze: | OpenAIRE |
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