Biophysical Characterization Of Macromolecules

National Institute Of Biomedical Imaging And Bioengineering, Bethesda

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Abstract

Experimental techniques employed are analytical ultracentrifugation, static and dynamic light scattering, field flow fractionation, isothermal titration microcalorimetry, differential scanning calorimetry, circular dichroism spectroscopy, steady-state anisotropy fluorescence, fluorescence spectroscopy, and surface plasmon resonance biosensing. Molecules studied in this project include anti-poly-gamma-D-glutamic acid antibodies, anthrax toxin binding to a series of anti-LF neutralizing antibodies, the adaptor proteins of T-cell activation SLP76 and ADAP, and glutamate receptors. A new method was developed for density contrast sedimentation velocity analytical ultracentrifugation, with the goal to allow the measurement of particle density in polydisperse samples. Also, we developed and verified an approach for the prediction of protein refractive indices. Finally, we have created a visual protocol for colleagues to accomplish ultracentrifugal cell assembly.