Method and apparatus for performing peptide digestion on a microfluidic device
First Claim
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1. A method of processing a protein sample in a microfluidic device, the method comprising the steps of:
- providing a microfluidic device comprising a sample reservoir, a waste reservoir, a plurality of parallel channels connecting the sample reservoir to the waste reservoir, and a plurality of reagent reservoirs that introduce reagents at various points along each of the plurality of parallel channels, placing immobilized trypsin in a portion of each of the plurality of channels, placing the reagents required to carry out reduction and alkylation, and quenching into the reagent reservoirs, applying a single negative pressure driving force to the waste reservoir that generates a continuous flow of fluid from the sample reservoir, through the plurality of channels, to the waste reservoir, wherein the flow sequentially exposes the fluid to the reduction, alkylation, and quenching reagents before the fluid flow through the immobilized trypsin, and collecting fluid in the waste reservoir.
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Abstract
The invention provides methods and apparatuses that allow a protein sample to undergo reduction, alkylation, and digestion in a continuous flow process carried out within a microfluidic device. Methods and apparatuses in accordance with the invention can be employed as part of an automated proteomics analysis carried out in an integrated proteomics system.
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Citations
1 Claim
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1. A method of processing a protein sample in a microfluidic device, the method comprising the steps of:
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providing a microfluidic device comprising a sample reservoir, a waste reservoir, a plurality of parallel channels connecting the sample reservoir to the waste reservoir, and a plurality of reagent reservoirs that introduce reagents at various points along each of the plurality of parallel channels, placing immobilized trypsin in a portion of each of the plurality of channels, placing the reagents required to carry out reduction and alkylation, and quenching into the reagent reservoirs, applying a single negative pressure driving force to the waste reservoir that generates a continuous flow of fluid from the sample reservoir, through the plurality of channels, to the waste reservoir, wherein the flow sequentially exposes the fluid to the reduction, alkylation, and quenching reagents before the fluid flow through the immobilized trypsin, and collecting fluid in the waste reservoir.
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Specification