Author/Authors :
Koreck?، نويسنده , , Lucie and B??lkov?، نويسنده , , Zuzana and Holèapek، نويسنده , , Michal and Kr?lovsk?، نويسنده , , Josef and Bene?، نويسنده , , Milan and Lenfeld، نويسنده , , Ji??? and Minc، نويسنده , , Nicolas and Cecal، نويسنده , , Roxana and Viovy، نويسنده , , Jean-Louis and Przybylski، نويسنده , , Michael، نويسنده ,
Abstract :
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.
