Ovalbumin Dotaz Zobrazit nápovědu
Specific allergen immunotherapy is frequently associated with adverse reactions. Several strategies are being developed to reduce the allergenicity while maintaining the therapeutic benefits. Peptide immunotherapy is one such approach. Methods for the simple and rapid identification of immunogenic epitopes of allergens (i.e. allergenic epitopes) are ongoing and could potentially lead to peptide-based vaccines. An epitope extraction technique, based on biofunctionalized magnetic microspheres self-organized under a magnetic field in a channel of a simple microfluidic device fabricated from polydimethylsiloxane, was applied in the isolation and identification of prospective allergenic epitopes. Similarly to chromatographic column separations, the easily replaceable plug of self-organized beads in the channel benefits especially from an even larger surface-to-volume ratio and an enhanced interaction of the surfaces with passing samples. Ovalbumin, the major protein of egg white and a typical representative of food allergens, was selected as the model molecule. Highly resistant ovalbumin was at first efficiently digested by a magnetic proteolytic reactor with trypsin treated with l-1-tosylamido-2-phenylethyl chloromethyl ketone and the second step, i.e. capture of allergenic epitopes from the mixture of peptides, was performed by a magnetic immunoaffinity carrier with orientedly immobilized rabbit anti-ovalbumin IgG molecules. Captured peptides were released with 0.05% trifluoroacetic acid. The elution fractions were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The peptide fragment of ovalbumin HIATNAVLFFGR (m/z: 1345.75, position: 371-382) was identified as a relevant allergenic epitope in this way. Such a microfluidic magnetic force-based epitope extraction technique applied in the epitope mapping of ovalbumin has the potential to be a significant step towards developing safe and cost-effective epitope-based vaccines.
- MeSH
- alergeny chemie imunologie MeSH
- epitopy analýza MeSH
- financování organizované MeSH
- hmotnostní spektrometrie MeSH
- imunomagnetická separace metody MeSH
- mapování epitopu metody MeSH
- mikrofluidní analytické techniky metody MeSH
- mikrosféry MeSH
- ovalbumin chemie imunologie MeSH
- potravinová alergie MeSH
- vakcíny MeSH
- MeSH
- adjuvancia imunologická MeSH
- Bacillus imunologie MeSH
- finanční podpora výzkumu jako téma MeSH
- imunizace metody MeSH
- myši MeSH
- ovalbumin aplikace a dávkování MeSH
- respirační sliznice sekrece MeSH
- transportní proteiny terapeutické užití MeSH
- vakcíny konjugované aplikace a dávkování MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- MeSH
- Escherichia coli MeSH
- exprese genu MeSH
- glutathiontransferasa MeSH
- ovalbumin MeSH
- proteinové inženýrství MeSH
- Publikační typ
- srovnávací studie MeSH
- MeSH
- citalopram farmakologie MeSH
- experimenty na zvířatech MeSH
- financování organizované MeSH
- inhibitory fosfodiesteras aplikace a dávkování diagnostické užití MeSH
- kašel farmakoterapie MeSH
- morčata MeSH
- nemoci dýchací soustavy farmakoterapie MeSH
- ovalbumin diagnostické užití škodlivé účinky MeSH
- purinony farmakologie MeSH
- tetrazoly farmakologie MeSH
- vinca alkaloidy farmakologie MeSH
- zvířata MeSH
- Check Tag
- morčata MeSH
- zvířata MeSH
- Publikační typ
- abstrakty MeSH
