DNA Aptamer for Molecular Recognition and Protein Expression Profiling

Aptamer has attracted much attention in both scientific community and pharmaceutical industry in the past few years. Its high affinity and specificity to target molecules have render aptamer with wide applications in both diagnostic and therapeutic fields. We demonstrate the molecular recognition capability of aptamers by conjugating them to resin beads and packing them in an HPLC column for affinity purification of Taq DNA polymerase. The concept of screening aptamers for a multitude of proteins is demonstrated by using protein extract of E. coli in a multiplex SELEX selection process. To reduce the wide dynamic range of protein concentration, the bacterial cell lysate was fractionated by two-dimensional chromatography. An aptamer microarray was constructed by using randomly selected 384 aptamer probes to illustrate the capability of aptamer microarray in profiling protein expression and in identifying differentially expressed genes. The aptamer microarray successfully discriminated two different bacterial species and identified a protein which was over-expressed when the E. coli was treated with ampicillin.

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