An analytical method to control the surface density and stability of DNA-gold nanoparticles for an optimized biosensor

DNA functionalized gold nanoparticles (DNA-AuNPs) have shown great potential for biosensing as they combine the excellent optical properties of gold nanoparticles and the molecular recognition function of DNA. Since the DNA density determines the assay performance and the stability of the conjugate, a precise control of the surface density of DNA-AuNP is crucial for an optimized biosensor. Here we report a simple assay for quantifying multiple unlabeled DNAs on AuNPs. The assay relies on potassium cyanide (KCN) to first dissolve the AuNPs, which then releases surface bound DNA for quantification through a double-stranded DNA dye. Using this analytical quantification method, we investigated several strategies to control the surface density of DNA-AuNPs. Besides the precise control of DNA density, the stability of DNA-AuNPs after conjugation is also important in developing a biosensor with optimal performance. Without proper storing conditions, DNA-AuNPs are unstable and aggregate over time. To overcome this problem, we developed a long-term storage solution to ensure the stability and quality of DNA-AuNPs after conjugation which would benefit any DNA-AuNP-based biosensor.