Rapid Generation of High-affinity Aptamers for Quantification of Protein Biomarkers

The generation of genomic and proteomic data with the goal of biomarker discovery for disease diagnostics or prognosis is a powerful approach. However, biomarkers identified from a typically small sample size must be validated on a large population. A limiting factor is the availability of high-quality reagents for the quantification of the biomarkers in a complex medium such as serum. The assay must be technically simple and amenable to high-throughput procedures. Screening of recombinant antibody libraries is a powerful method for the generation of affinity reagents. However, “affinity maturation” techniques required to produce high affinity antibodies is a relatively slow procedure as it requires library isolation, diversification, and retransformation back into cells. Radiation Monitoring Devices, Inc. proposes the high-throughput generation of nucleic acids-based aptamer affinity reagents. We introduce a novel technique for generating aptamers with high affinity and specificity. In addition, we will utilize a screening technique that does not require sample modification with labels or separation of the bound/unbound aptamers. This will increase the number of clones for which we are able to obtain binding data. Finally, because selection occurs outside of cells, the entire procedure is rapid including all maturation steps for improving the binding characteristics.