Responsive Hydrogel Binding Matrix for Dual Signal Amplification in Fluorescence Affinity Biosensors and Peptide Microarrays
Language English Country United States Media print-electronic
Document type Journal Article
- Keywords
- biomarkers, click chemistry, microarrays, pNIPAAm, peptide, plasmon-enhanced fluorescence, serotesting, thermoresponsive hydrogel,
- MeSH
- Acrylic Resins chemistry MeSH
- Biosensing Techniques methods MeSH
- Fluorescence MeSH
- Hydrogels chemistry metabolism MeSH
- Immunoglobulin G analysis immunology MeSH
- Epstein-Barr Virus Infections diagnosis immunology metabolism virology MeSH
- Humans MeSH
- Peptide Fragments immunology metabolism MeSH
- Polymers chemistry MeSH
- Epstein-Barr Virus Nuclear Antigens immunology MeSH
- Herpesvirus 4, Human immunology isolation & purification MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Acrylic Resins MeSH
- Hydrogels MeSH
- Immunoglobulin G MeSH
- Peptide Fragments MeSH
- poly-N-isopropylacrylamide MeSH Browser
- Polymers MeSH
- Epstein-Barr Virus Nuclear Antigens MeSH
A combined approach to signal enhancement in fluorescence affinity biosensors and assays is reported. It is based on the compaction of specifically captured target molecules at the sensor surface followed by optical probing with a tightly confined surface plasmon (SP) field. This concept is utilized by using a thermoresponsive hydrogel (HG) binding matrix that is prepared from a terpolymer derived from poly(N-isopropylacrylamide) (pNIPAAm) and attached to a metallic sensor surface. Epi-illumination fluorescence and SP-enhanced total internal reflection fluorescence readouts of affinity binding events are performed to spatially interrogate the fluorescent signal in the direction parallel and perpendicular to the sensor surface. The pNIPAAm-based HG binding matrix is arranged in arrays of sensing spots and employed for the specific detection of human IgG antibodies against the Epstein-Barr virus (EBV). The detection is performed in diluted human plasma or with isolated human IgG by using a set of peptide ligands mapping the epitope of the EBV nuclear antigen. Alkyne-terminated peptides were covalently coupled to the pNIPAAm-based HG carrying azide moieties. Importantly, using such low-molecular-weight ligands allowed preserving the thermoresponsive properties of the pNIPAAm-based architecture, which was not possible for amine coupling of regular antibodies that have a higher molecular weight.
FZU Institute of Physics Czech Academy of Sciences Na Slovance 2 Prague 182 21 Czech Republic
References provided by Crossref.org
Plasmon-Enhanced Multiphoton Polymer Crosslinking for Selective Modification of Plasmonic Hotspots
Plasmonic nanomaterials with responsive polymer hydrogels for sensing and actuation
Rapid Actuation of Thermo-Responsive Polymer Networks: Investigation of the Transition Kinetics
