Peptide epitope-imprinted polymer microarrays for selective protein recognition
- We introduce a practically generic approach for the generation of epitope-imprinted polymer-based microarrays for protein recognition on surface plasmon resonance imaging (SPRi) chips. The SPRi platform allows the subsequent rapid screening of target binding kinetics in a multiplexed and label-free manner. The versatility of such microarrays, both as synthetic and screening platform, is demonstrated through developing highly affine molecularly imprinted polymers (MIPs) for the recognition of the receptor binding domain (RBD) of SARS-CoV-2 spike protein. A characteristic nonapeptide GFNCYFPLQ from the RBD and other control peptides were microspotted onto gold SPRi chips followed by the electrosynthesis of a polyscopoletin nanofilm to generate in one step MIP arrays. A single chip screening of essential synthesis parameters, including the surface density of the template peptide and its sequence led to MIPs with dissociation constants (K-D) in the lower nanomolar range for RBD, which exceeds the affinity of RBD for its natural target,We introduce a practically generic approach for the generation of epitope-imprinted polymer-based microarrays for protein recognition on surface plasmon resonance imaging (SPRi) chips. The SPRi platform allows the subsequent rapid screening of target binding kinetics in a multiplexed and label-free manner. The versatility of such microarrays, both as synthetic and screening platform, is demonstrated through developing highly affine molecularly imprinted polymers (MIPs) for the recognition of the receptor binding domain (RBD) of SARS-CoV-2 spike protein. A characteristic nonapeptide GFNCYFPLQ from the RBD and other control peptides were microspotted onto gold SPRi chips followed by the electrosynthesis of a polyscopoletin nanofilm to generate in one step MIP arrays. A single chip screening of essential synthesis parameters, including the surface density of the template peptide and its sequence led to MIPs with dissociation constants (K-D) in the lower nanomolar range for RBD, which exceeds the affinity of RBD for its natural target, angiotensin-convertase 2 enzyme. Remarkably, the same MIPs bound SARS-CoV-2 virus like particles with even higher affinity along with excellent discrimination of influenza A (H3N2) virus. While MIPs prepared with a truncated heptapeptide template GFNCYFP showed only a slightly decreased affinity for RBD, a single mismatch in the amino acid sequence of the template, i.e. the substitution of the central cysteine with a serine, fully suppressed the RBD binding.…
Author details: | Zsófia BognárORCiD, Eszter SupalaORCiD, Aysu YarmanORCiDGND, Xiaorong ZhangGND, Frank Fabian BierORCiDGND, Frieder W. SchellerORCiDGND, Róbert E. GyurcsanyiORCiD |
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DOI: | https://doi.org/10.1039/d1sc04502d |
ISSN: | 2041-6539 |
Pubmed ID: | https://pubmed.ncbi.nlm.nih.gov/35222909 |
Title of parent work (English): | Chemical science / RSC, Royal Society of Chemistry |
Subtitle (English): | application for SARS-CoV-2 RBD protein |
Publisher: | Royal Society of Chemistry |
Place of publishing: | Cambridge |
Publication type: | Article |
Language: | English |
Date of first publication: | 2021/11/23 |
Publication year: | 2022 |
Release date: | 2024/01/05 |
Volume: | 13 |
Issue: | 5 |
Number of pages: | 8 |
First page: | 1263 |
Last Page: | 1269 |
Funding institution: | NRDI Fund (TKP2020 IES, BME-IENAT) of the Ministry for Innovation and Technology; Germany's Excellence Strategy [EXC 2008-390540038-UniSysCat]; German Ministry of Education and Research (BMBF)Federal Ministry of Education & Research (BMBF) [01DH20018] |
Organizational units: | Mathematisch-Naturwissenschaftliche Fakultät / Institut für Biochemie und Biologie |
DDC classification: | 5 Naturwissenschaften und Mathematik / 54 Chemie / 540 Chemie und zugeordnete Wissenschaften |
Peer review: | Referiert |
Publishing method: | Open Access / Hybrid Open-Access |
License (English): | Creative Commons - Namensnennung - Nicht-kommerziell - Weitergabe unter gleichen Bedingungen 3.0 Unported |