Methods and devices for protein assays
View Patent ↗Methods and devices for protein assays based on Edman degradation in microfluidic channels are disclosed herein. As disclosed, the cleaved amino acid residues may be immobilized in an array format and identified by detectable labels, such as antibodies, which specifically bind given amino acid residues. Alternatively, the antibodies are immobilized in an array format and the cleaved amino acids are labeled identified by being bound by the antibodies in the array.
1. An assay comprising
subjecting a protein to consecutive coupling and cleaving steps to result in a plurality of cleaved amino acid residues in a microfluidic device,
contacting each cleaved amino acid residue with a plurality of antibodies, which each antibody specifically binds a given amino acid,
identifying each cleaved amino acid residue by the antibody bound thereto, and
identifying the location of each cleaved amino acid residue in the sequence of the protein.
2. The assay of claim 1 , wherein each antibody is conjugated to a detectable label.
3. The assay of claim 1 , wherein the cleaved amino acid residues are immobilized in a microchannel in an array based on its time of cleavage in the consecutive coupling and cleaving steps.
4. The assay of claim 1 , wherein the antibodies are immobilized in an array in parallel microchannels.
5. The assay of claim 3 , which further comprises obtaining at least 19 parallel microchannels.
6. The assay of claim 3 , wherein the each antibody is conjugated to a distinguishable label and the antibodies are contacted with the cleaved amino acid residues at substantially the same time in one step and then each cleaved amino acid residue and its location are identified.
7. The assay of claim 3 , wherein each antibody is sequentially contacted with the cleaved amino acid residues and the cleaved amino acid residues and their locations are identified by deduction after contact with each antibody.
8. The assay of claim 5 , wherein each antibody is contacted with the plurality of cleaved amino acid residues in one parallel microchannel and then each amino acid residue and its location are identified.
9. The assay of claim 4 , wherein each cleaved amino acid residue is contacted with the plurality of antibodies in one parallel microchannel and then each amino acid residue and its location are identified.
10. The assay of claim 1 , wherein the cleaved amino acid residues and their locations are identified by sequentially detecting each cleaved amino acid by chromatography.
11. A microfluidic device comprising
at least one microchannel having an amino acid residue bound to an antibody that specifically binds the amino acid residue and said antibody is conjugated to a detectable label.
12. The microfluidic device of claim 11 , and further comprising one or more parallel microchannels.