Protocol for measuring anti-fentanyl antibodies in mouse serum by enzyme-linked immunosorbent assay

Jacob A. Koster; Alejandra Prevost-Reilly; Dheeraj Soni; Aisling Kelly; Kristina Walker; Elizabeth B. Norton; Ofer Levy; Colin N. Haile; David J. Dowling

STAR Protocols (Dec 2024)

Protocol for measuring anti-fentanyl antibodies in mouse serum by enzyme-linked immunosorbent assay

Jacob A. Koster,
Alejandra Prevost-Reilly,
Dheeraj Soni,
Aisling Kelly,
Kristina Walker,
Elizabeth B. Norton,
Ofer Levy,
Colin N. Haile,
David J. Dowling

Affiliations

Jacob A. Koster: Precision Vaccines Program, Department of Pediatrics, Boston Children’s Hospital, Boston, MA 02115, USA
Alejandra Prevost-Reilly: Precision Vaccines Program, Department of Pediatrics, Boston Children’s Hospital, Boston, MA 02115, USA
Dheeraj Soni: Precision Vaccines Program, Department of Pediatrics, Boston Children’s Hospital, Boston, MA 02115, USA; Department of Pediatrics, Harvard Medical School, Boston, MA 02115, USA; Global Investigative Toxicology, Preclinical Safety, Sanofi, Cambridge, MA, USA
Aisling Kelly: Precision Vaccines Program, Department of Pediatrics, Boston Children’s Hospital, Boston, MA 02115, USA
Kristina Walker: Precision Vaccines Program, Department of Pediatrics, Boston Children’s Hospital, Boston, MA 02115, USA
Elizabeth B. Norton: Department of Microbiology and Immunology, Tulane University School of Medicine, New Orleans, LA 70112, USA
Ofer Levy: Precision Vaccines Program, Department of Pediatrics, Boston Children’s Hospital, Boston, MA 02115, USA; Department of Pediatrics, Harvard Medical School, Boston, MA 02115, USA; Broad Institute of MIT & Harvard, Cambridge, MA 02142, USA
Colin N. Haile: Department of Psychology/TIMES, University of Houston, Houston, TX 77204, USA
David J. Dowling: Precision Vaccines Program, Department of Pediatrics, Boston Children’s Hospital, Boston, MA 02115, USA; Department of Pediatrics, Harvard Medical School, Boston, MA 02115, USA; Corresponding author

Journal volume & issue: Vol. 5, no. 4
p. 103402

Abstract

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Summary: 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.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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Abstract

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