Fabrication and characterization of conjugate nano-biological systems interfacing metallic nanostructures on solid supports with immobilized biomolecules is reported. The entire sequence of relevant experimental steps is described, involving the fabrication of nanostructured substrates using electron beam lithography, immobilization of biomolecules on the substrates, and their characterization utilizing surface-enhanced Raman spectroscopy (SERS). Three different designs of nano-biological systems are employed, including protein A, glucose binding protein, and a dopamine binding DNA aptamer. In the latter two cases, the binding of respective ligands, D-glucose and dopamine, is also included. The three kinds of biomolecules are immobilized on nanostructured substrates by different methods, and the results of SERS imaging are reported. The capabilities of SERS to detect vibrational modes from surface-immobilized proteins, as well as to capture the protein-ligand and aptamer-ligand binding are demonstrated. The results also illustrate the influence of the surface nanostructure geometry, biomolecules immobilization strategy, Raman activity of the molecules and presence or absence of the ligand binding on the SERS spectra acquired.
Copyright © 2015 Journal of Visualized Experiments
bio-functionalized surfaces, proteins, aptamers, molecular recognition, nanostructures, electron beam lithography, surface-enhanced Raman spectroscopy
Peters, Robert F., Luis Gutierrez-Rivera, Steven K. Dew, and Maria Stepanova. "Surface enhanced Raman spectroscopy detection of biomolecules using EBL fabricated nanostructured substrates." Journal of Visualized Experiments 97 (2015): e52712.
Journal of Visualized Experiments