Quantitation of Albumin in Serum Using “Turn-on” Fluorescent Probe with Aggregation-Enhanced Emission Characteristics
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Abstract
An aggregation-enhanced emission active luminogen named as sodium 4,4'4″-(3,4-diphenyl-1H-pyrrole-1,2,5-triyl)tribenzoate (DP-TPPNa) with propeller construction was synthesized and developed as a "turn on" fluorescent probe for in situ quantitation of albumin in blood serum. The DP-TPPNa fluorescence intensity was linearly correlated with the concentration of two serum albumins, bovine serum albumin (BSA) and human serum albumin (HSA), in pure PBS buffer in the ranges of 2.18-70 and 1.68-100 μg/mL, respectively. The detection limits were as low as 2.18 μg/mL for BSA and 1.68 μg/mL for HSA. The response time of fluorescence to serum albumin (SA) was very short (below 6 s), which achieved real-time detection. It also showed high selectivity to SA because other components in serum barely interfere with the detection of DP-TPPNa to SA, enabling in situ quantitative detection of SA without isolation from serum. DP-TPPNa was successfully applied for the quantitative detection of BSA in fetal bovine serum. The mechanism of fluorescent turn-on behavior was elucidated utilizing an unfolding process induced by guanidine hydrochloride, which revealed a capture process via selective hydrophobic interaction and hydrogen bonding between luminogen and SA.
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