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 into a single volume 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 are not split during the sample preparation process and where common purification methods can be used for purifying multiple analytes.
1. A method for simultaneously co-purifying and concentrating nucleic acid and protein targets, the method comprising:
obtaining a sample comprising nucleic acids and protein;
applying said sample to a hand-held apparatus comprising a portable device wherein the portable device comprises a gel-based isotachophoresis separation unit and a detection unit for the detection of an analyte selected from the group consisting of nucleic acid and protein,
wherein the detection unit comprises disposable cartridges,
subjecting said sample to a gel-based isotachophoresis on a cartridge of the portable device with the proviso that the isotachophoresis is not by the method of capillary electrophoresis,
wherein the gel-based isotachophoresis comprises buffer formulations comprising Bis-tris MES solution in the gel, a bicine/NaOH cathode buffer and a Bis-tris/acetic acid solution as an anode buffer; and
simultaneously co-purifying and concentrating nucleic acid and protein targets into a single volume in buffer by application of a voltage wherein an amount of charge imparted to the nucleic acid and protein targets causes simultaneous concentration of the nucleic acids and protein targets in a single volume; and
obtaining the concentrated nucleic acid and protein targets from the hand-held apparatus; and
detecting the presence of the nucleic acid and protein targets.
2. The method of claim 1 wherein the step of detecting the presence of protein targets comprises a method selected from the group consisting of immunoassay, protein sequencing, mass spectrometry, functional assays, non-antibody ligand binding, gels, and/or combinations thereof; and the step of detecting the presence of nucleic acid targets comprises a method 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 of the presence of the nucleic acid and protein targets is performed in a single output.