Protein classification by autofluorescence spectral shape analysis using machine learning

Label-free visualization of unfolding and crosslinking mediated protein aggregation in nonenzymatically glycated proteins

“…Therefore, the maximum Fluorescence Resonance Energy Transfer (FRET) from Tyr to Trp residues 22,28 makes the Tyr-specific peak weakly visible in the dUV-AF spectra of Hb compared to HSA. 28 Also, irrespective of the donor (Tyr) and acceptor (Trp) numbers, the proteins produce either resolved or unresolved dUV-AF spectra, 29 which can be attributed to the different discrete states (or classes) and spatial arrangement of Trp residues in proteins that determine their spectral shapes and emission maxima. 30 However, the more resolved peaks of Tyr and Trp in HSA help in assessing the unfolding through the Tyr/Trp intensity ratio 31 (Fig.…”

“…A high-resolution charge-coupled device (CCD) spectrometer (QEPro, Ocean Optics, Inc., USA) was used to record the autofluorescence spectra of glycated proteins. 22 On the emission side, a 300 nm long-pass filter (FF01-300/LP, Semrock, USA) was used to block the excitation light from the LED, allowing only the autofluorescence spectra of glycated proteins. The glycated protein samples (incubated for three days) were loaded into an in-house designed quartz multi-well plate (with 36 quartz wells), and the dUV-AF spectra were then recorded at room temperature (25 °C).…”