Development of Human Serum Albumin Selective Fluorescent Probe Using Thieno[3,2-b]pyridine-5(4H)-one Fluorophore Derivatives

The level of human serum albumin (HSA) in biological fluids is a key health indicator and its quantitative determination has great clinical importance. In this study, we developed a selective and sensitive fluorescent HSA probe by fluorescence-based high-throughput screening of a set of fluorescent thieno[3,2-b]pyridine-5(4H)-one derivatives against major plasma proteins: HSA, bovine serum albumin (BSA), globulin, fibrinogen, and transferrin. The fluorophore chosen finally (4 in Scheme 1) showed noticeable fluorescence enhancement in the presence of HSA (160-fold increase), and it exhibited rapid response, high sensitivity (detection limit 8 nM), and the ability to clearly distinguish HSA from BSA in pH 9 buffer condition. Moreover, the probe could be applicable to detect trace amounts of HSA in an artificial urine sample; further, it might be applied to the determination of the HSA concentration in complex biological samples for pre-clinical diagnosis.

Download Full-text

Chemometrics-assisted multi-instrumental techniques for investigation of interactions of dapagliflozin with normal and glycated human serum albumin: Application to exploiting second-order advantage for determination of glycated human serum albumin as a biomarker for controlling diabetes

Microchemical Journal ◽

10.1016/j.microc.2021.106313 ◽

2021 ◽

Vol 167 ◽

pp. 106313

Author(s):

Asma Mohamady ◽

Mohsen Shahlaei ◽

Vali Akbari ◽

Hector C. Goicoechea ◽

Ali R. Jalalvand

Keyword(s):

Human Serum Albumin ◽

Serum Albumin ◽

Human Serum ◽

Second Order

Download Full-text

Simultaneous Determination of Human Serum Albumin and Low-Molecular-Weight Thiols after Derivatization with Monobromobimane

Molecules ◽

10.3390/molecules26113321 ◽

2021 ◽

Vol 26 (11) ◽

pp. 3321

Author(s):

Katarzyna Kurpet ◽

Rafał Głowacki ◽

Grażyna Chwatko

Keyword(s):

Molecular Weight ◽

Human Serum Albumin ◽

Serum Albumin ◽

Human Serum ◽

Simultaneous Determination ◽

Reversed Phase ◽

Fluorescence Labeling ◽

Detector Response ◽

Low Molecular Weight

Biothiols are extremely powerful antioxidants that protect cells against the effects of oxidative stress. They are also considered relevant disease biomarkers, specifically risk factors for cardiovascular disease. In this paper, a new procedure for the simultaneous determination of human serum albumin and low-molecular-weight thiols in plasma is described. The method is based on the pre-column derivatization of analytes with a thiol-specific fluorescence labeling reagent, monobromobimane, followed by separation and quantification through reversed-phase high-performance liquid chromatography with fluorescence detection (excitation, 378 nm; emission, 492 nm). Prior to the derivatization step, the oxidized thiols are converted to their reduced forms by reductive cleavage with sodium borohydride. Linearity in the detector response for total thiols was observed in the following ranges: 1.76–30.0 mg mL−1 for human serum albumin, 0.29–5.0 nmol mL−1 for α-lipoic acid, 1.16–35 nmol mL−1 for glutathione, 9.83–450.0 nmol mL−1 for cysteine, 0.55–40.0 nmol mL−1 for homocysteine, 0.34–50.0 nmol mL−1 for N-acetyl-L-cysteine, and 1.45–45.0 nmol mL−1 for cysteinylglycine. Recovery values of 85.16–119.48% were recorded for all the analytes. The developed method is sensitive, repeatable, and linear within the expected ranges of total thiols. The devised procedure can be applied to plasma samples to monitor biochemical processes in various pathophysiological states.

Download Full-text