Plate-Adherent Nanosubstrate for Improved ELISA of Small Molecules: A Proof of Concept Study

Enzyme-linked immunosorbent assay (ELISA) is a widely used technique for detecting and quantifying target analytes in clinical and research laboratories. One of the main drawbacks of ELISA is the involvement of multiple washing steps that desorbs the capture antigen/antibody off the polystyrene plat...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2020-08, Vol.92 (16), p.10952-10956
Main Authors: Ghoshdastidar, Shreya, Gangula, Abilash, Kainth, Jagjot, Saranathan, Sandhya, Elangovan, Ashwath, Afrasiabi, Zahra, Hainsworth, Dean P, Upendran, Anandhi, Kannan, Raghuraman
Format: Article
Language:English
Subjects:
8-Hydroxy-2'-Deoxyguanosine - chemistry
8-Hydroxy-2'-Deoxyguanosine - urine
8-Hydroxydeoxyguanosine
Adsorption
Analytical chemistry
Animals
Antibodies
Antigens
Cattle
Chemistry
Deoxyguanosine
Enzyme-linked immunosorbent assay
Enzyme-Linked Immunosorbent Assay - methods
Gelatin
Gelatin - chemistry
Laboratories
Nanoparticles
Nanoparticles - chemistry
Polystyrene
Polystyrene resins
Proof of Concept Study
Serum Albumin, Bovine - chemistry
Stability analysis
Substrates
Target detection
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Summary:Enzyme-linked immunosorbent assay (ELISA) is a widely used technique for detecting and quantifying target analytes in clinical and research laboratories. One of the main drawbacks of ELISA is the involvement of multiple washing steps that desorbs the capture antigen/antibody off the polystyrene plate, thereby producing inconsistent and erroneous data. To overcome the problem of desorption, we hypothesized that gelatin nanoparticles (GelNP) could serve as a “plate-adherent” substrate to irreversibly adhere the capture antigen/antibody of interest. We tested our hypothesis using GelNP-based substrate (Gel–BSA–OHG) to adhere 8-hydroxy-2′-deoxyguanosine (8-OHdG) to the polystyrene plate and assayed this molecule using the ELISA technique. The stability and ELISA performance of Gel–BSA–OHG was evaluated in comparison to the conventional substrate (BSA–OHG). Importantly, the Gel–BSA–OHG substrate was found to be more wash-resistant and consequently resulted in improved sensitivity, accuracy, and precision in the ELISA analysis of 8-OHdG. Finally, the scope of Gel–BSA–OHG substrate-based ELISA for clinical application was demonstrated by validating its ability to detect 8-OHdG in an artificial urine sample with high specificity.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.0c01441