Microarray based Raman spectroscopic detection with gold nanoparticle probes.
A microarray approach based on surface-enhanced Raman spectroscopic (SERS) was developed for detection of spotted peptide, peptide-protein or protein-antibody interaction. The procedure involves the attachment of peptide-capped gold nanoparticles followed by silver deposition for signal enhancement. The attachment of the gold nanoparticles is achieved by standard avidin-biotin chemistry. The well-known biomolecular recognition pairs, IgG/protein A and biotin/avidin, were used to demonstrate proof-of-concept of the SERS assay. Detection limits of 10 and 100 fg per microarray spot were obtained respectively for the peptide and protein arrays. For the protein in solution, a limit of 0.1 microg/mL is reported. Furthermore, enzyme activity of the kinase (PKA) is also detected with high specificity for an established peptide substrate (kemptide) on the microarray spots.
Li
Tao
T
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, China.
Guo
Liping
L
Wang
Zhenxin
Z
D004867:Equipment Design; D019544:Equipment Failure Analysis; D006046:Gold / Q000737:chemistry*; D015336:Molecular Probe Techniques / Q000295:instrumentation*; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure*; D010316:Particle Size; D040081:Protein Array Analysis / Q000295:instrumentation* / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013059:Spectrum Analysis, Raman / Q000295:instrumentation* / Q000379:methods
D006046:Gold