Purification and concentration of proteins and DNA from a complex sample using isotachophoresis and a device to perform the purification
A method of simultaneously co-purifying and concentrating nucleic acid and protein targets is described. The method includes automation of the entire sample preparation process, performed by having an analyst add a sample into a device that performs all of the steps necessary to prepare a sample for analysis. The method provides for samples that are not split during the sample preparation process and where common purification methods can be used for purifying multiple analytes.
1. A method of simultaneously separating and concentrating nucleic acid and protein targets, the steps comprising:
(i) obtaining a sample comprising a nucleic acid and a protein;
(ii) adding said sample to a gel electrophoresis unit, said gel electrophoresis includes a gel box with a negative electrode side and a positive electrode side, the negative electrode side being filled with a first buffer comprising 2-Hydroxy-N-(tris(hydroxymethyl) methyl)-3-aminopropanesulfonic acid buffer and the positive electrode side being filled with a second buffer being different than said first buffer;
(iii) subsequent to step (ii), subjecting said sample to isotachophoresis, said subjecting step comprising a step of applying a voltage to said gel electrophoresis unit; and
(iii) subsequent to step (iii), detecting the presence of the nucleic acid and the protein.
2. The method of claim 1 wherein the detecting the presence of protein is selected from the group consisting of immunoassay, protein sequencing, mass spectrometry, functional assays, non-antibody ligand binding, aptamers, gels, and/or combinations thereof; and the detecting the presence of nucleic acid is selected from the group consisting of PCR, isothermal amplification methods, hybridization reactions, microarrays, protein-DNA binding, mass spectrometry, gels, and/or combinations thereof.
3. The method of claim 1 wherein the detecting the presence of protein is selected from the group consisting of immunoassay, protein sequencing, mass spectrometry, functional assays, non-antibody ligand binding, aptamers, gels, and/or combinations thereof; and the detecting the presence of nucleic acid is selected from the group consisting of PCR, isothermal amplification methods, hybridization reactions, microarrays, protein-DNA binding, mass spectrometry, gels, and/or combinations thereof, and wherein the detecting the presence of nucleic acid and protein are performed in a single output.
4. The method of claim 1 wherein subjecting said sample to isotachophoresis comprises simultaneously purifying and concentrating both the nucleic acid and the protein by driving the nucleic acid toward a positive electrode and driving the protein toward a negative electrode responsive to application of a voltage.
5. The method of claim 4 wherein the sample is subjected to isotachophoresis on a cartridge of a handheld device.
6. The method of claim 1 wherein subjecting said sample to isotachophoresis comprises simultaneously purifying and concentrating both the nucleic acid and the protein, the purifying being accomplished by driving the nucleic acid toward a positive electrode and driving the protein toward a negative electrode responsive to application of a voltage, the concentrating being simultaneously accomplished based on the first and second buffers causing separation of different nucleic acids or proteins while the different nucleic acids or proteins move in opposite directions so that different nucleic acids are separated from each other or different proteins are separated from each other while moving in respective opposite directions responsive to the application of a voltage based on respective different charges associated with the different nucleic acids or proteins in the buffer.
7. The method of claim 1 wherein the gel box contains a solidified gel matrix including a top surface, said negative electrode side being filled with said first buffer to a first buffer fill level being equal to or below said top surface of the solidified gel matrix and said positive electrode side being filled with said second buffer to a second buffer fill level being equal to or below said top surface of the solidified gel matrix.