Protein discrimination based on DNA induced perylene probe self-assembly

Abstract

The development of a simple and effective method for the highly sensitive and selective discrimination of proteins is a subject of enormous interest. Herein, we report the construction of a novel fluorescence detection method based on a perylene probe for the highly efficient discrimination of multiple proteins. Single-stranded DNA (ssDNA) could induce aggregation of the perylene probe which caused quenching of probe fluorescence. After the addition of a protein, the protein could interact with the ssDNA-probe assembly complex with "turn-on" or further "turn-off" fluorescence response. A sensor array was designed based on the above phenomena which could realize the successful discrimination of proteins with 100% accuracy of cross validation. Nine representative proteins were successfully recognized. Moreover, it was observed that a protein could induce characteristic effect on the DNA-probe assembly with varying pH of assay buffer. Thus, different proteins showed unique fluorescence response towards assay buffers having different pH values. The assay buffer pH was then utilized as a sensing channel. Based on Linear Discriminant Analysis (LDA) nine proteins were successfully discriminated at the nanomolar concentration with 100% accuracy of cross validation. Furthermore, the sensor array also demonstrated differentiation of the nine proteins regardless of their concentration. The developed sensor array could also detect the proteins with great precision in human urine sample at a quite low concentration, which suggests its practical applicability for analysis of biological fluids.

Keywords: Array; Fluorescence; Perylene probe; Protein discrimination; Self-assembly; ssDNA.