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

doi:10.1016/j.xpro.2024.103402

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

STAR Protoc. 2024 Dec 20;5(4):103402. doi: 10.1016/j.xpro.2024.103402. Epub 2024 Dec 6.

Authors

Jacob A Koster¹, Alejandra Prevost-Reilly¹, Dheeraj Soni², Aisling Kelly¹, Kristina Walker¹, Elizabeth B Norton³, Ofer Levy⁴, Colin N Haile⁵, David J Dowling⁶

Affiliations

¹ Precision Vaccines Program, Department of Pediatrics, Boston Children's Hospital, Boston, MA 02115, USA.
² 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.
³ Department of Microbiology and Immunology, Tulane University School of Medicine, New Orleans, LA 70112, USA.
⁴ 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.
⁵ Department of Psychology/TIMES, University of Houston, Houston, TX 77204, USA.
⁶ Precision Vaccines Program, Department of Pediatrics, Boston Children's Hospital, Boston, MA 02115, USA; Department of Pediatrics, Harvard Medical School, Boston, MA 02115, USA. Electronic address: [email protected].

Abstract

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.

Keywords: antibody; cell biology; immunology.

MeSH terms

Animals
Antibodies* / blood
Antibodies* / immunology
Enzyme-Linked Immunosorbent Assay* / methods
Fentanyl* / blood
Fentanyl* / immunology
Mice
Mice, Inbred BALB C*
Mice, Inbred C57BL*

Substances

Fentanyl
Antibodies