A polystyrene ELISA plate (EP) modified with a thin film based on gold nanoseeds (AuSDs) assembled onto polydopamine (PDA) is proposed. The nanodecorated film (PDA@AuSD) allows to evaluate the polyphenols antioxidant capacity (AOC) through a colorimetric approach based on a seed-mediated growth strategy. Polyphenols, in the presence of the nanodecorated (PDA@AuSD) surfaces are able to drive an increase in size of the AuSDs according to their AOC; this produces an increase of the localized surface plasmon resonance (LSPR; maximum at λ ~ 550 nm) that is taken as analytical signal. The PDA@AuSD EP manufacturing shows good intraplates repeatability (RSD ≤ 6.6%, n = 96 wells) and interplates reproducibility (RSD ≤ 7.4%, n = 748 wells), resulting stable for 1 year. The AuSDs growth kinetic has been studied using 11 polyphenols belonging to different chemical classes and 4 different food samples. The PDA@AuSD film is able to return quantitative information on the AOC of food polyphenols. Good repeatability (RSD ≤ 5.7%, n = 12 EP wells) and reproducibility (RSD ≤ 8.1%, n = 12 EP wells) was achieved, with acceptable linear correlation coefficients (R ≥ 0.990) and useful limits of detection (LODs ≤ 2.5 10 mol L). The samples analyzed with the PDA@AuSD device have been successfully ordered according to their AOC in agreement with conventional optical methods. The PDA@AuSD plate allows multiple measurements (96 wells per EP) with a one-step strategy, overcoming the limitations related to the use of colloidal nanoparticles; in addition, since absorbance is measured after washing, it is not affected by sample color or turbidity. Graphical abstract Schematic representation of ELISA plate (EP) modified with polydopamine (PDA) film decorated with gold nanoseeds (AuSD). The colorimetric assay, to evaluate the antioxidant capacity, is based on the AuSD growth mediated by polyphenols, resulting in absorbance increase at 550 nm (ΔAbs), which is employed as analytical signal.
Download full-text PDF |
Source |
---|---|
http://dx.doi.org/10.1007/s00604-020-04228-4 | DOI Listing |
J Extracell Biol
January 2025
RoseBio Milano Italy.
Current state-of-the-art tools for analysing extracellular vesicles (EVs) offer either highly sensitive but unidimensional bulk measurements of EV components, or high-resolution multiparametric single-particle analyses which lack standardization and appropriate reference materials. This limits the accuracy of the assessment of marker abundance and overall marker distribution amongst individual EVs, and finally, the understanding of true EV heterogeneity. In this study, we aimed to define the standardized operating procedures and reference material for fluorescent characterization of EVs with two commonly used EV analytical platforms-nanoparticle tracking analysis (NTA) and nano-flow cytometry (nFCM).
View Article and Find Full Text PDFPathogens
December 2024
Centro Nacional de Investigación Disciplinaria en Salud Animal e Inocuidad, Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias, Ciudad de México 05110, Mexico.
(LPMV) is the etiologic agent of blue eye disease (BED), which affects pigs of all ages, and it has been endemic in central Mexico since the 1980s. To date, no disease control program has been established. Therefore, there is a need for a serological diagnostic method with high sensitivity and specificity.
View Article and Find Full Text PDFJ Appl Microbiol
January 2025
G.B. Elyakov Pacific Institute of Bioorganic Chemistry FEB RAS; 690022 Vladivostok, Russia.
Aims: The aim of this study was to evaluate the antioxidant and anti-inflammatory effects of marine fungal cerebroside flavuside B (FlaB) on Staphylococcus aureus-infected keratinocytes in in vitro skin wounds and to identify FlaB targets in bacterial and human cells.
Methods And Results: A combination of ELISA, plate spectrofluorimetry, and flow cytometry with fluorescence dye staining, scratch assay, and real-time cell imaging techniques was used to investigate the effects of FlaB on S. aureus-infected HaCaT keratinocytes.
Sci Rep
January 2025
Department of Veterinary Parasitology, U. P. Pandit Deen Dayal Upadhyaya Pashu Chikitsa Vigyan Vishwavidyalaya Evam Go Anusandhan Sansthan (DUVASU), Mathura, 281001, India.
This study is the first to conduct a sero-surveillance of Bovine Tropical Theileriosis (BTT) caused by the protozoan parasite Theileria annulata (T. annulata) using a recombinant Tams1 protein-based dot-ELISA in cattle, and to compare its efficacy with plate-ELISA, single PCR, nested PCR, and blood microscopy. The goal was to identify the most effective method for the early and accurate detection of theileriosis, which significantly impacts livestock through reduced milk yield and increased mortality.
View Article and Find Full Text PDFPol J Vet Sci
December 2024
Pasteur Institute of Novi Sad, Hajduk Veljkova 1, 21101 Novi Sad, Serbia.
Constant antigenic changes, new variants and easy transmission of SARS-CoV-2 virus should acquire greater zoonotic attention and need to remain alert. In this retrospective study the aim was to analyze seropositivity to SARS-CoV-2 in dogs by commercial ELISA. The Virus neutralization test (VNT) was modified for the purpose of confirmation of SARS-CoV-2 antibodies in ELISA-positive dog sera.
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!