scholarly journals Protein-templated copper nanoclusters for fluorimetric determination of human serum albumin

2021 ◽  
Vol 188 (4) ◽  
Author(s):  
Mariagrazia Lettieri ◽  
Pasquale Palladino ◽  
Simona Scarano ◽  
Maria Minunni
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Fluorescent Detection ◽  
Fluorimetric Determination ◽  
Copper Nanoclusters ◽  
Optical Reporters ◽  
The Stability ◽  
Serum And Urine

AbstractCopper nanoclusters (CuNCs) are attractive for their unique optical properties, providing sensitive fluorescent detection of several kinds of targets even in complex matrices. Their ability in growing on suitable protein and nucleic acid templates make CuNCs efficient optical reporters to be exploited in bioanalysis. In this work, we report the specific and sensitive determination of human serum albumin (HSA) in human serum (HS) and urine via CuNCs fluorescence. HSA is the most abundant protein in plasma, and plays a key role in the early diagnosis of serious pathological conditions such as albuminuria and albuminemia. Recently, HSA has become clinically central also as a biomarker to assess severity, progression, and prognosis of various cancers. We report the controlled and reproducible growth of CuNCs directly on the target analyte, HSA, which results in a fine dose-dependent fluorescent emission at 405 nm. The protocol is optimized in water, and then applied to serum and urine specimens, without matrix pretreatment. The method linearly responds within the whole concentration of clinical interest, with a sensitivity of 1.8 ± 0.1 × 10−3 g L−1 and 0.62 ± 0.03 × 10−3 g L−1 in serum and urine, respectively, and excellent reproducibility (CVav% ca. 3% for both). The assay is designed to have a single protocol working for both matrices, with recovery of 95% (HS) and 96% (urine). The stability of the fluorescence after CuNCs formation was tested over 3 days, displaying good results (yet higher in urine than in serum). Graphical abstract

scholarly journals Protein-Templated Copper Nanoclusters: Versatile Platform for Label-Free Detection of Albumin

Proceedings ◽  
2020 ◽  
Vol 60 (1) ◽  
pp. 55
Author(s):  
Mariagrazia Lettieri ◽  
Pasquale Palladino ◽  
Simona Scarano ◽  
Maria Minunni
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Accurate Determination ◽  
Quantitative Detection ◽  
Metal Nanoclusters ◽  
Copper Nanoclusters ◽  
Wide Range ◽  
Label Free Detection

The outstanding properties of metal nanoclusters, stabilized with different scaffolds, i.e., proteins, nucleic acids, polymers and dendrimers, enable their application in a wide range of fields. The recent advances in the fabrication and synthesis of nanoclusters have revolutionized the design of biosensors, leading to significant improvements in the selective and sensitive determination of several targets. In particular, in recent years, copper nanoclusters (CuNCs) have attracted more attention mainly for their unique fluorescent properties, as well as their large Stokes shifts, low toxicity, and high biocompatibility. The high-photoluminescent features of CuNCs facilitate highly sensitive target detection even in complex biological matrices. For these reasons, in this work, we exploited the specific template-targeted CuNCs’ growth for the sensitive and accurate determination of human serum albumin (HSA) in urine and human serum. HSA is the most abundant protein in plasma, acting as a carrier for many key biological molecules such as hormones, fatty acids and steroids, and it contributes to the maintenance of the oncotic blood pressure. The concentration of HSA in body fluids greatly influences the state of health of the patients. Taking into account these considerations, the quantitative detection of human serum albumin plays a key role in the early diagnosis of serious pathological conditions such as albuminuria and albuminemia. Here, we present a CuNCs-based assay in which copper nanoclusters were used as fluorescent signal indicators to detect serum albumin in a complex biological matrix.

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

Molecules ◽  
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.

scholarly journals The Influence of Oxidative Stress on Serum Albumin Structure as a Carrier of Selected Diazaphenothiazine with Potential Anticancer Activity

2021 ◽  
Vol 14 (3) ◽  
pp. 285
Author(s):  
Małgorzata Maciążek-Jurczyk ◽  
Beata Morak-Młodawska ◽  
Małgorzata Jeleń ◽  
Wiktoria Kopeć ◽  
Agnieszka Szkudlarek ◽  
...  
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Anticancer Activity ◽  
Drug Distribution ◽  
Cd Spectroscopy ◽  
Drug Binding ◽  
Protein Activity ◽  
Chloramine T ◽  
The Stability

Albumin is one of the most important proteins in human blood. Among its multiple functions, drug binding is crucial in terms of drug distribution in human body. This protein undergoes many modifications that are certain to influence protein activity and affect its structure. One such reaction is albumin oxidation. Chloramine T is a strong oxidant. Solutions of human serum albumin, both non-modified and modified by chloramine T, were examined with the use of fluorescence, absorption and circular dichroism (CD) spectroscopy. 10H-3,6-diazaphenothiazine (DAPT) has anticancer activity and it has been studied for the first time in terms of binding with human serum albumin—its potential as a transporting protein. Using fluorescence spectroscopy, in the presence of dansylated amino acids, dansyl-l-glutamine (dGlu), dansyl-l-proline (dPro), DAPT binding with two main albumin sites—in subdomain IIA and IIIA—has been evaluated. Based on the conducted data, in order to measure the stability of DAPT complexes with human (HSA) and oxidized (oHSA) serum albumin, association constant (Ka) for ligand-HSA and ligand-oHSA complexes were calculated. It has been presumed that oxidation is not an important issue in terms of 10H-3,6-diazaphenothiazine binding to albumin. It means that the distribution of this substance is similar regardless of changes in albumin structure caused by oxidation, natural occurring in the organism.

Comparison of the stability of Y-90-, Lu-177- and Ga-68- labeled human serum albumin microspheres (DOTA-HSAM)

2010 ◽  
Vol 37 (8) ◽  
pp. 861-867 ◽  
Author(s):  
Gerd Wunderlich ◽  
Eik Schiller ◽  
Ralf Bergmann ◽  
Hans-Jürgen Pietzsch
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Albumin Microspheres ◽  
The Stability
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