Microfluidic system for proteome analysis
First Claim
1. An microfluidic device comprising a substrate having an at least one sample holding channel for receiving a substantially purified polypeptide, wherein said at least one sample holding channel collects said substantially purified polypeptide in order to enable a chemical or physical process to occur.
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Abstract
The invention provides a microfluidic system and method to rapidly analyze large numbers of compounds or complex mixtures of compounds, particularly, low abundance cellular proteins involved in cell signaling pathways. In one aspect, an integrated microfluidic system comprises an upstream separation module (preferably, a multi-dimensional separation device), a microfluidic device for on-device protein digestion of substantially separated proteins received from the upstream separation module, a downstream separation module for separating digestion products of said proteins, a peptide analysis module and a processor for determining the amino acid sequence of said proteins. Preferably, the system comprises an interfacing microfluidic device between the downstream separation module and the peptide analysis module.
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Citations
51 Claims
- 1. An microfluidic device comprising a substrate having an at least one sample holding channel for receiving a substantially purified polypeptide, wherein said at least one sample holding channel collects said substantially purified polypeptide in order to enable a chemical or physical process to occur.
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20. A method for protein digestion comprising:
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(a) delivering a sample to an at least one sample holding channel in a microfluidic device; and
(b) exposing said sample to a protease for a sufficient period of time to obtain a desired amount of a digestion product. - View Dependent Claims (21, 22, 23)
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- 24. An integrated microfluidic system for proteome analysis comprising a microfluidic module having a microfluidic device wherein said microfluidic module is in communication with an upstream separation module for providing a substantially purified polypeptide to an at least one sample holding channel of said microfluidic device.
- 32. An integrated microfluidic system for proteome analysis comprising a microfluidic module having a microfluidic device wherein said microfluidic module is in communication with a downstream separation module for separating a plurality of digestion products wherein said digestion products result from a digestion reaction taking place on said microfluidic device.
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47. A method for proteome analysis digestion comprising:
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(a) providing an integrated microfluidic system for proteome analysis;
(b) delivering a sample comprising a plurality of cellular polypeptides to an upstream separation module of said integrated microfluidic system and obtaining a substantially purified polypeptide;
(c) delivering said substantially purified polypeptide to a microfluidic module of said integrated microfluidic system wherein said microfluidic module comprises a protease, exposing said polypeptide to said protease for a period of time and under a set of conditions sufficient to substantially digest said polypeptide, thereby producing an at least one digestion product;
(d) transporting said digestion product to a downstream separation module of said system, and obtaining a substantially separated digestion product; and
(e) determining an amino acid sequence of said digestion product and assembling said sequences to generate a sequence for said substantially purified polypeptide. - View Dependent Claims (48, 49, 50, 51)
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Specification