Target Peptide Detection and Measurement Using a Capacitive Immunoprobe Biosensor
Published: April 30, 2024
Abstract
Source: Kluge, N. et al., Time-Resolved In Vivo Measurement of Neuropeptide Dynamics by Capacitive Immunoprobe in Porcine Heart. J. Vis. Exp.(2022).
This video demonstrates a method for detecting and quantifying peptide transmitters using a capacitive immunoprobe biosensor. The functionalized electrode, coated with antibodies, interacts with peptides of interest, leading to a signal change that allows precise measurement and detection of peptides.
Protocol
1. Capacitive immunoprobe fabrication and functionalization Cut a 25 cm length of perfluoroalkoxy (PFA)-coated platinum wire (see Table of Materials) and strip approximately 5 mm of PFA coating from one end using a scalpel, being careful not to cut into the platinum wire. Insert the stripped end of the platinum wire into a gold-plated 1 mm male connector pin and crimp the connector pin teeth around the stripped end of the platinum wire using needle-nose pliers …
Disclosures
The authors have nothing to disclose.
Materials
| AgCl disc electrode | Warner Instruments (Holliston, MA) | 64-1307 | |
| Anti-NPY monoclonal antibody | Abcam, (Cambridge, MA) | ab112473 | |
| Custom multichannel amplifier/ 1 MΩ feedback resistor multichannel headstage | NPI Electronic, (Tamm, Germany) | NA | Based on NPI VA-10M multichannel amplifier |
| Dopamine HCl | Sigma Aldrich (St. Louis, MO) | H8502-10G | |
| Gold-plated male connector pin | AMP-TE Connectivity (Amplimite) | 6-66506-1 | |
| HEKA LIH 8+8 analog-to-digital/digital-to-analog device | HEKA Elektronik, (Holliston, MA) | NA | |
| Igor Pro data acquisition software, v. 7.08 | WaveMetrics, (Lake Oswego, OR) | Software driving command potential and data acquisition was custom written | |
| Masterflex L/S Standard Digital peristaltic pump | Cole Palmer, (Vernon Hills, IL) | ||
| PFA-coated platinum wire | A-M Systems, (Sequim, WA) | 773000 | 0.005" bare diameter, 0.008" coated diameter |
| Silicone elastomer | World Precision Instruments (Sarasota, FL) | SYLG184 | |
| Synthetic porcine NPY peptide | Bachem (Torrance, CA) | 4011654 |
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Cite This Article
Target Peptide Detection and Measurement Using a Capacitive Immunoprobe Biosensor. J. Vis. Exp. (Pending Publication), e22128, doi: (2024).