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Immunization of mice with the self-peptide ACBP coupled to keyhole limpet hemocyanin

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2022

STAR Protoc. 2022 Jan 13;3(1):101095. doi: 10.1016/j.xpro.2021.101095. eCollection 2022 Mar 18.

Immunization of mice with the self-peptide ACBP coupled to keyhole limpet hemocyanin

Léa Montégut, Hui Chen, José Manuel Bravo-San Pedro, Omar Motiño, Isabelle Martins, Guido Kroemer

Centre de Recherche des Cordeliers, Equipe labellisée par la Ligue Contre le Cancer, Université de Paris, Sorbonne Université, INSERM U1138, Institut Universitaire de France, Paris, France. Metabolomics and Cell Biology Platforms, Institut Gustave Roussy, Villejuif, France. Université Paris Saclay, Faculty of Medicine Kremlin Bicêtre, Le Kremlin Bicêtre, France. Complutense University of Madrid, Faculty of Medicine, Department Section of Physiology, Madrid, Spain. Pôle de Biologie, Hôpital Européen Georges Pompidou, AP-HP, Paris, France.

Service type: Knockout mice

Abstract

Keyhole limpet hemocyanin (KLH) is a glycosylated multi-subunit metalloprotein that elicits a strong nonspecific immune activation, thus inducing both cellular and humoral immune responses. The exceptional immunogenicity of this protein can be leveraged to vaccinate mice against self-antigens that otherwise would not induce an autoimmune response. This protocol describes the covalent conjugation of KLH with acyl-coenzyme A-binding protein (ACBP), the autovaccination of mice with ACBP-KLH conjugate together with a potent adjuvant, and the detection of the produced anti-ACBP autoantibodies. For complete details on the use and execution of this profile, please refer to Bravo-San Pedro et al. (2019c).

Keywords: Immunology; Metabolism; Model Organisms; Molecular Biology.

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