Author(s): Carneiro, Mariana C. C. G. ; Rodrigues, Lígia R. ; Moreira, Felismina ; Sales, M. Goreti F.
Date: 2024
Persistent ID: http://hdl.handle.net/10400.22/29471
Origin: Repositório Científico do Instituto Politécnico do Porto
Subject(s): Molecularly imprinted polymer; Biomimetic enzyme-linked immunosorbent assay; Point-of-care; Cellulose paper; Cancer antigen 15–3; Protein biomarker
The combination of molecularly-imprinted polymers (MIPs) on paper substrates for capturing a cancer antigen 15–3 (CA15-3) with the traditional coloured transduction scheme of enzyme-linked immunosorbent assay (ELISA) using 3,3′,5,5′-tetramethylbenzidine (TMB), horseradish peroxidase (HRP) and H2O2 is presented here for the first time. Here, the paper surface was modified with a silane derivative containing an amine function that allows subsequent binding of 3-aminophenylboronic acid (3-APBA). The target protein, CA15-3, was then bound via the boronic groups of APBA. The empty space around CA15-3 was filled by polymerization of dopamine. Finally, the CA15-3 template was removed by breaking the imine function to create vacant sites for which CA15-3 has a high affinity. Binding of CA15-3 was detected by oxidation of TMB substrate by HRP in the presence of H2O2. The results showed that the MIP was able to selectively recognize CA15-3 within 3–500 U mL−1, even in complex samples. Quantitative data were extracted by analysing the colour coordinates of images captured with a smartphone using ImageJ. This selective behaviour was confirmed by comparison with a control material (non– polymer, NIP). Overall, the described approach has advantages over conventional ELISA, namely low cost and high stability, which is due to the use of MIP as a trapping element acting as a selective pre-concentration point of CA15-3. To our knowledge, this is the first biomimetic ELISA (B-ELISA) on paper substrate used for macromolecules such as proteins. We believe that this approach has the potential to be applied to other protein disease biomarkers, making it a suitable tool for screening glycoproteins at the point-of-care.
