Integrated Microreactor/Nanoelectrospray Emitter
Proteins broken-up into peptide components (using proteolytic enzymes) are injected into mass spectrometers for analysis using Nano-Electrospray Ionization (Nano-ESI) techniques. During capillary electrophoresis, one of the most favored techniques in the ionization-spray phase, some portions of peptides adsorb (stick) to the inner walls of the capillaries. This results in loss of expensive sample as well as in analytical inaccuracies. This technology describes a technique to immobilize the prote olytic enzymes such as trypsin on the inner walls of capillaries so that peptide adsorption is reduced and process efficiency is enhanced. Through the same, the new technique also provides for a high degree of integration amongst the proteolysis, sep aration and ionization phases since almost all protein can now be broken down and separated at the point of contact with the immobilized enzyme, obviating the requirement for a separate chromatographic separation phase. Other values of this technolog y are:
a) While protein digestion by trypsin in solution takes 18-24 hours, trypsin in immobilized form digests the protein in only 15 minutes even for diluted concentrations (100x), resulting in high digestion efficiency b) No trypsin autodigest ion products produced c) The sequence coverage of amino acids within the protein is as high as 90%
d) Sample requirement is minimal (~ 1 µL) e) Immobilization is done using covalent chemical bonding; easy cleaning and reusability possible
The technology can be employed in any scenario that requires protein analysis using mass spectrometry.
Categories: Genomics, Nanotechnology
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