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A biomimetic enzyme-linked immunosorbent assay (BELISA) for the analysis of gonadorelin by using molecularly imprinted polymer-coated microplates.
| Content Provider | Europe PMC |
|---|---|
| Author | Torrini, Francesca Caponi, Laura Bertolini, Andrea Palladino, Pasquale Cipolli, Francesca Saba, Alessandro Paolicchi, Aldo Scarano, Simona Minunni, Maria |
| Abstract | An original biomimetic enzyme-linked immunoassay (BELISA) to target the small peptide hormone gonadorelin is presented. This peptide has been recently listed among the substances banned in sports by the World Antidoping Agency (WADA) since its misuse by male athletes triggers testosterone increase. Hence, in response to this emerging issue in anti-doping controls, we proposed BELISA which involves the growth of a polynorepinephrine (PNE)–based molecularly imprinted polymer (MIP) directly on microwells. PNE, a polydopamine (PDA) analog, has recently displayed impressive performances when it was exploited for MIP preparation, giving even better results than PDA. Gonadorelin quantification was accomplished via a colorimetric indirect competitive bioassay involving the competition between biotinylated gonadorelin linked to the signal reporter and the unlabeled analyte. These compete for the same MIP binding sites resulting in an inverse correlation between gonadorelin concentration and the output color signal (λ = 450 nm). A detection limit of 277 pmol L−1 was achieved with very good reproducibility in standard solutions (avCV% = 4.07%) and in urine samples (avCV% = 5.24%). The selectivity of the assay resulted adequate for biological specimens and non-specific control peptides. In addition, the analytical figures of merit were successfully validated by mass spectrometry, the reference anti-doping benchtop platform for the analyte. BELISA was aimed to open real perspectives for PNE-based MIPs as alternatives to antibodies, especially when the target analyte is a poorly or non-immunogenic small molecule, such as gonadorelin.Graphical abstractBiomimetic enzyme-linked immunosorbent assay (BELISA)Supplementary InformationThe online version contains supplementary material available at 10.1007/s00216-021-03867-7. |
| ISSN | 16182642 |
| Journal | Analytical and Bioanalytical Chemistry |
| Volume Number | 414 |
| PubMed Central reference number | PMC9242967 |
| Issue Number | 18 |
| PubMed reference number | 35028691 |
| e-ISSN | 16182650 |
| DOI | 10.1007/s00216-021-03867-7 |
| Language | English |
| Publisher | Springer Berlin Heidelberg |
| Publisher Date | 2022-01-13 |
| Publisher Place | Berlin/Heidelberg |
| Access Restriction | Open |
| Rights License | Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. © The Author(s) 2022 |
| Subject Keyword | Molecularly imprinted polymers Antibody mimetics Polydopamine Polynorepinephrine Enzyme-linked immunosorbent assay Gonadotropin-releasing hormone |
| Content Type | Text |
| Resource Type | Article |
| Subject | Analytical Chemistry Biochemistry |
