RESUMO
A new strategy for encoding polypeptide libraries with photolabile tags is developed. The photoassisted assay, based on conditional release of encoding tags only from bound pairs, can differentiate between peptides which have minor differences in a form of post-translational modifications with epigenetic marks. The encoding strategy is fully compatible with automated peptide synthesis. The encoding pendants are compact and do not perturb potential binding interactions.
RESUMO
Externally sensitized electron-transfer fragmentation in dithiane PPG (photoremovable protecting group)-protected carbonyls is adopted for detection and amplification of molecular recognition events. The new methodology allows for detection of as low as 50 attomoles of avidin utilizing an imager based on a low sensitivity mass-produced consumer CCD camera. Numeric modelling is carried out to demonstrate the intrinsic limitations of 2D amplification on surfaces and the advantages of unconstrained amplification in a compartmentalized volume of spatially addressable 3D solutions.
Assuntos
Benzofenonas/química , Corantes Fluorescentes/química , Benzofenonas/síntese química , Sítios de Ligação , Simulação por Computador , Quinolizinas/química , Compostos de Enxofre/químicaRESUMO
Amplified fluorescence quenching methodology based on massive autocatalytic photo-unmasking of a dual function sensitizer-quencher is developed and adopted for photoassisted ultra-sensitive detection of molecular recognition events. The resulting binding assay, based on a molecular recognition-triggered photo-amplified cascade with concomitant decrease of fluorescence is validated with the biotin-avidin pair, achieving attomolar detection.