Working with others, we developed biospecific self-assembled monolayer (SAM) chips that increased, by more than an order of magnitude, the sensitivity, accuracy, and range of measurements made with surface plasmon resonance (SPR) instruments. We have extended the strategies developed for our biospecific nitrilotri-acetic acid (NTA) SAM chips to the development of a DNA presenting monolayer chip. This new chip should extend the sensing enhancements demonstrated by the NTA-SAM to the study of DNA-protein complex interactions. We will also explore how a highly ordered surface of known structure might be used to probe macromolecules of unknown structure. SPR determined kinetic measurements will be calibrated against solution based measurements obtained with fluorescence anisotropy. As with the development of the NTA-SAM, we will employ the power of genetics to assess the accuracy of our experimental surfaces. We have available arrays of well characterized proteins and variants thereof, whose activation potential can be measured in vivo, so that the relevance of in vitro measurements can be ascertained.
