Publication detail

Tryptic digestion using self-organized magnetic particles on chip.

Year: 2005

Type of publication: ostatní - přednáška nebo poster

Name of source: 18th International Symposium in Microscale Bioseparations

Publisher name: 18th International Symposium in Microscale Bioseparations

Place:

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Titles:

Language	Name	Abstract	Keywords
cze	Tryptické štěpení s použitím samouspořádaných magnetických partikulí v čipu	Tryptické štěpení s použitím samouspořádaných magnetických partikulí v čipu	trypsin; microfluidic device; kinetic measurement
eng	Tryptic digestion using self-organized magnetic particles on chip.	A microsystem was developed for fast inline microfluidic digestion of proteins with off-chip analysis. Trypsin catalyses the proteolysis of protein, amide, or ester bonds at the carboxyl linkage of L-lysine or L-arginine residues. This enzyme was grafted on carboxylated magnetic particles by carbodiimide (EDC and S-NHS) activation. Grafting was optimized, in order to control the magnetic bead tendency to aggregate, non-specific sorption, proteolytic activity and shelf life. These particles were used as a microcolumn, packed by self-organization in a strong and oriented magnetic field gradient inside a microchannel prepared in (PDMS) by soft lithography. Model protein recombinant human growth hormone was digested in the microfluidic channel and efficiency was measured as a function of flow rates and residence time by SDS-PAGE, CE or MALDI MS. Efficient and highly specific digestion in 3 min was demonstrated with sequence covering over 48% and peptide map comparable to digestion in solution.	trypsin reactor; microfluidic device; kinetic measurement

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