Screening of compounds interacting with HIV-1 proteinase using optical biosensor technology.

摘要: A high-resolution optical biosensor assay for screening of low-molecular-weight compounds, using an immobilized protein target, has been developed. HIV-1 proteinase was on the sensor surface by direct amine coupling and a variety inhibitors noninteracting reference drugs were applied to in continuous flow buffer. The procedure did not require intrinsic reporter groups, substrates, inhibitors, or other ligands detection. By protein, signal could be corrected relatively large background caused differences dimethyl sulfoxide concentration between running sample buffers. Substances binding with high affinity (Ki nM range) required efficient regeneration washing injection system cycles get consistent results. Analysis simplified report points, extracted during both association dissociation phases, simple graphical display data. optimized correctly distinguish from compounds randomized series, indicate their interaction kinetics, reveal artifacts due nonspecific signals, incomplete regeneration, carryover. method is expected generally applicable secondary compound libraries proteins as targets.