Covalent binding of streptavidin on gold magnetic nanoparticles for bead array fabrication.
In this study, we report the chemical synthesis and functionalization of streptavidin coated gold magnetic nanoparticles (GMNPs) and the immobilization of single-stranded biotinylated oligonucleotides onto these particles. By using covalent interaction or physical adsorption, two kinds of streptavidin coated GMNPs (SA-GMNPs) were prepared. The quantity and stability of streptavidin bound to the GMNPs using different methods were determined by UV-Vis spectrometer. The results indicated that by physical absorption the GMNPs can capture more streptavidin, the SA-GMNPs with either physical adsorption or covalent reaction were both stable in PBS buffer. In contrast, SA-GMNPs with covalent reaction was stable in SDS buffer, while most of the SA-GMNPs by physical adsorption would be eluted from the particles in SDS buffer. Therefore, the SA-GMNPs by covalent immobilization were more suitable for fabrication of bead array. To evaluate the binding efficiency and capacity of DNA on SA-GMNPs, the capture of biotinylated oligonucleotide or PCR products on SA-GMNPs at different concentrations were examined. A magnetic beads array was fabricated by immobilizing DNA-MNPs complexes onto a glass slide using a magnetic field. The synthesized DNA targets with different concentrations were detected with a detection limit of approximately 0.05 nM, indicating the potential application of this MNPs array to high-throughput DNA detection.
State Key Laboratory of Bioelectronics (Chien-Shiung Wu Laboratory), Department of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China.
Song
S
Li
Hongna
H
Liu
Nongyue
N
He
D000327:Adsorption; D001710:Biotin / Q000737:chemistry / Q000378:metabolism; D006046:Gold / Q000737:chemistry*; D055767:Immobilized Proteins / Q000737:chemistry* / Q000378:metabolism; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D008863:Microspheres; D036103:Nanotechnology / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D009841:Oligonucleotides / Q000032:analysis; D055550:Protein Stability; D012680:Sensitivity and Specificity; D019809:Streptavidin / Q000737:chemistry* / Q000378:metabolism
D055767:Immobilized Proteins; D058185:Magnetite Nanoparticles; D009841:Oligonucleotides; D001710:Biotin; D006046:Gold; D019809:Streptavidin