Protocol for measuring anti-fentanyl antibodies in mouse serum by enzyme-linked immunosorbent assay
Enzyme-linked immunosorbent assay (ELISA) offers an effective, inexpensive, and reliable approach for the analysis of humoral immune responses. Here, we describe a protocol for measuring anti-fentanyl antibodies generated by the immunization of mice with novel opioid vaccine candidates. We describe steps for coating BSA-fentanyl antigen and standard wells. We then detail procedures for preparing buffers and performing assays. This protocol is adaptable for use with either BALB/c or C57BL/6J mice strains and can measure antibody isotypes to elucidate class switching. For complete details on the use and execution of this protocol, please refer to Stone et al.<sup>1</sup>.
Authors
- Aisling Kelly (ORCID: https://orcid.org/0009-0006-7872-6719)
- David J. Dowling (ORCID: https://orcid.org/0000-0003-1095-6156)
- Jacob A. Koster (ORCID: https://orcid.org/0000-0002-4645-6308)
- Colin N. Haile (ORCID: https://orcid.org/0000-0001-8293-7291)
- Ofer Levy (ORCID: https://orcid.org/0000-0002-5859-1945)
- Elizabeth B. Norton (ORCID: https://orcid.org/0000-0002-8426-3515)
- Alejandra Prevost-Reilly (ORCID: https://orcid.org/0000-0003-4360-4763)
- Dheeraj Soni (ORCID: https://orcid.org/0000-0002-5555-2051)
- Kristina Walker
Institutions
- Broad Institute (US)
- Tulane University (US)
- Boston Children's Hospital (US)
- University of Houston (US)
Publication Details
- Journal
- STAR Protocols
- Published
- 2024-12-01
- DOI
- https://doi.org/10.1016/j.xpro.2024.103402
- Primary Topic
- Anesthesia and Sedative Agents
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- National Institutes of Health