2015-034 – Two Dimensional Fluorescence Difference Spectroscopy (2D FDS) for Characterization of Nanoparticles Interactions with Biomolecules

Nanoparticles are being developed as a platform to support a number of technologies. For example, in drug development antibodies, proteins, vaccines, and other beneficial compounds are attached to the nanoparticles for transport into the body and greatly increase the effectiveness of treatments. To ensure that patients are receiving the treatments, it is important to have a method to confirm the quality and reproducibility of nanoparticles, and even more importantly to validate their attachment to drug, antibody, environmental contaminants or other molecules. Current processes for this are somewhat non-quantitative and non-specific. For example, visualization by electron microscopy and changes in the electrical charge at the surface or size of the particle are methods used to verify attachment of compounds to the nanoparticles. However, these methods are not sensitive enough and are certainly not amenable to high throughput screening. In order to address these issues, researchers at Kansas State University have developed a novel concept for concurrently scanning both excitation and emission wavelengths while also categorizing the light intensity of fluorescent nanoparticles. Here light beams of pre-specified wavelengths are sequentially passed through the sample and a 2-dimensional spectrum is built for the fluorescent excitation and emission pattern. Under these conditions most nanoparticles emit a characteristic fluorescent signature or fingerprint which is material-dependent existing at the maximal excitation/emission intersect. Moreover, this 2-dimensional fluorescence difference spectroscopy (2-D FDS) and the 2-D spot intrinsic to each nanoparticle undergoes a shift when the nanoparticle is bound by the biomolecule or substance to which it is attached. This allows for the researcher to confirm the attached compound or vaccine presence and concentration by checking the excitation or emission wavelengths. Each wavelength is indicative of a different phenomenon pertaining to the nanoparticle and its attachments. The ability to check for both excitation and emission wavelengths at the same time provides a higher level of quality assurance that the vaccine, antibody, or protein has attached to the nanoparticle and is ready for delivery to the body. Caronda Moore caronda@ksu.edu 785-532-1366