Rapid Sub-nanomolar Protein Determination in Serum using Electropolymerized Molecularly Imprinted Polymers (E-MIPs)

Stephen, Andrei N, Dennison, Sarah Rachel orcid iconORCID: 0000-0003-4863-9607, Holden, Mark orcid iconORCID: 0000-0003-3060-7615 and Reddy, Subrayal M orcid iconORCID: 0000-0002-7362-184X (2023) Rapid Sub-nanomolar Protein Determination in Serum using Electropolymerized Molecularly Imprinted Polymers (E-MIPs). The Analyst . ISSN 0003-2654

[thumbnail of VOR]
PDF (VOR) - Published Version
Available under License Creative Commons Attribution Non-commercial.


Official URL: https://doi.org/10.1039/D3AN01498C


Rapid detection of biologicals is important for a range of applications such as medical screening and diagnostics. Antibodies are typically employed for biosensing with high sensitivity and selectivity but can take months to prepare. Here, we investigate electropolymerized molecularly imprinted polymers (E-MIPs), which are produced in minutes as alternative-antibody rapid biosensors for the selective recognition of model proteins bovine haemoglobin (BHb) and bovine serum albumin (BSA). We evaluated two disposable screen-printed electrodes (SPE) designated AT-Au and BT-Au based on their different annealing temperatures. E-MIPs for BHb demonstrated an imprinting factor of 146:1 at 1nM and 12:1 at 0.1nM, showing high effectiveness of E-MIPs compared to their control non-imprinted polymers. The BHb imprinted E-MIP, when tested against BSA as a non-target protein, gave a selectivity factor of 6:1 for BHb. Sensor sensitivity directly depended on the nature of the SPE, with AT-Au SPE demonstrating limits of detection in the sub-micromolar range typically achieved for MIPs, while BT-Au SPE exhibited sensitivity in the sub-nanomolar range for target protein. We attribute this to differences in electrode surface area between AT-Au and BT-Au SPEs. The E-MIPs were also tested in calf serum as a model biological medium. The BT-Au SPE MIPs detected the presence of target protein in < 10 min with an LOD of 50 pM and LOQ of 100 pM, suggesting their suitability for protein determination in serum with minimal sample preparation. Using electrochemical impedance spectroscopy, we determine equilibrium dissociation constants (KD) for E-MIPs using the Hill-Langmuir adsorption model. KD of BHb E-MIP was determined to be 0.86 ± 0.11nM.

Repository Staff Only: item control page