High-throughput detection of protein???protein and protein???DNA interactions via photonic crystal biosensors: applications to apoptosis inducing factor

摘要

High-throughput screening (HTS) has played an integral role in the discovery of leadmolecules that eventually are transformed into the therapies of tomorrow. Contributing towardsthe success of any screening campaign is the target choice, chemical library size and diversity, aswell as the technology utilized in screening. Photonic crystal (PC) biosensors are a relativelynew technology whose potential is only beginning to be realized. PC biosensors are a ???labelfree???technology compatible with standard high-throughput techniques. Macromolecularinteractions including protein???small molecule, protein???nucleic acid, protein???protein, andwhole cell interactions can be measured using PC biosensors. Presented herein is the firstapplication of PC biosensors to a protein???nucleic acid interaction and a simplified protein???protein interaction platform. The protein???nucleic acid interaction targeted is the apoptosisinducing factor (AIF)???DNA interaction, which is critical for the mediation of caspaseindependentcell death in neurodegenerative disease and stroke. Protein???protein interactionswere targeted by modifying the PC biosensor surface for detection of the His6 tag, which iswidely employed in protein purification.