Simultaneous Purification and Fractionation of Nucleic Acids and Proteins from Complex Samples Using Bidirectional Isotachophoresis

We report on our efforts to create an on-chip system to simultaneously purify and fractionate nucleic acids and proteins from complex samples using isotachophoresis (ITP). We have developed this technique to simultaneously extract extracellular DNA and proteins from human blood serum samples and deliver these to two separate output reservoirs on a chip. The purified DNA is compatible with quantitative polymerase chain reaction (qPCR), and proteins can be extracted so as to exclude albumin, the most abundant protein in serum. We describe significant remaining challenges in making this bidirectional method a robust and efficient technique. These challenges include managing channel surface adsorption of proteins, identifying the cause of observed reductions in low molecular weight proteins, and dealing with nonspecific binding of proteins and DNA.