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

doi:10.1016/j.xpro.2021.101095

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

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

Authors

Léa Montégut^{1

2

3}, Hui Chen^{1

2

3}, José Manuel Bravo-San Pedro^{1

2

4}, Omar Motiño^{1

2}, Isabelle Martins^{1

2}, Guido Kroemer^{1

2

5}

Affiliations

¹ 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.
² Metabolomics and Cell Biology Platforms, Institut Gustave Roussy, Villejuif, France.
³ Université Paris Saclay, Faculty of Medicine Kremlin Bicêtre, Le Kremlin Bicêtre, France.
⁴ Complutense University of Madrid, Faculty of Medicine, Department Section of Physiology, Madrid, Spain.
⁵ Pôle de Biologie, Hôpital Européen Georges Pompidou, AP-HP, Paris, France.

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Carrier Proteins*
Hemocyanins
Immunization*
Mice
Peptides
Vaccination

Substances

Carrier Proteins
Peptides
Hemocyanins
keyhole-limpet hemocyanin