A novel substrate-inspired fluorescent probe to monitor native albumin in human plasma and living cells.

Human albumin (HA) displays crucial roles in maintaining health and fighting diseases. Accurate determination of native HA in plasma or non-plasma samples are of immense significance in both basic research and clinical practice. Herein, a novel ratiometric two-photon fluorescent probe (N-butyl-4-(4-phenyl-benzoyloxy) 1,8-naphthalimide, BPBN) has been designed and developed for highly selective and sensitive sensing the enzymatic activities of HA, on the basis of its unique pseudo-esterase feature. BPBN exhibits excellent selectivity, high sensitivity and good reactivity under physiological conditions. As an enzymatic activity-based probe, BPBN can distinguish between native HA and denatured HA, while the currently used dye-binding method cannot. The probe has been successfully applied to measure native HA in plasma samples and the secreted HA in the hepatocyte culture supernatant. BPBN has also been used for two-photon imaging of HA reabsorption in living renal cells, and the results demonstrate that this probe exhibits good cell permeability, low cytotoxicity and high imaging resolution. All these findings suggest that BPBN can be reliably used for the highly selective and sensitive detection of native HA in complex biological samples, as well as for exploring HA-associated biological processes and the physiological functions of native HA in living cells.