Utilization of newly developed immobilized enzyme reactors for preparation and study of immunoglobulin G fragments

0088382 - ÚMCH 2008 RIV NL eng J - Článek v odborném periodiku
Korecká, L. - Bílková, Z. - Holčapek, M. - Královský, J. - Beneš, Milan J. - Lenfeld, Jiří - Minc, N. - Cecal, R. - Viovy, J.-L. - Przybylski, M.
Utilization of newly developed immobilized enzyme reactors for preparation and study of immunoglobulin G fragments.
[Využití nově vyvinutých imobilizovaných enzymových reakcí pro přípravu a studii fragmentů imunoglobulinu G.]
Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences. Roč. 808, č. 1 (2004), s. 15-24. ISSN 1570-0232. E-ISSN 1873-376X.
[International Symposium on Polymer Design for BioSeparation and Nanobiotechnology /8./. Compiegne, 27.11.2003-29.11.2003]
Grant ostatní: GA ČR(CZ) GA203/02/0023
Program: GA
Výzkumný záměr: CEZ:AV0Z4050913
Klíčová slova: immobilized enzyme reactors * immunoglobulin G
Kód oboru RIV: CE - Biochemie
Impakt faktor: 2.176, rok: 2004

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.

Využití nově vyvinutých imobilizovaných enzymových reakcí pro přípravu a studii fragmentů imunoglobulinu G.
Trvalý link: http://hdl.handle.net/11104/0149929