Utilization of newly developed immobilized enzyme reactors for preparation and study of immunoglobulin G fragments
Jazyk angličtina Země Nizozemsko Médium print
Typ dokumentu časopisecké články, práce podpořená grantem
PubMed
15236681
DOI
10.1016/j.jchromb.2004.04.035
PII: S1570023204003988
Knihovny.cz E-zdroje
- MeSH
- bioreaktory * MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- enzymy metabolismus MeSH
- hmotnostní spektrometrie MeSH
- imunoglobulin G metabolismus MeSH
- imunoglobuliny - fragmenty chemie metabolismus MeSH
- molekulární sekvence - údaje MeSH
- sekvence aminokyselin MeSH
- spektroskopie infračervená s Fourierovou transformací MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- enzymy MeSH
- imunoglobulin G MeSH
- imunoglobuliny - fragmenty MeSH
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.
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