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.
STAR Protoc. 2022 Jan 13;3(1):101095. doi: 10.1016/j.xpro.2021.101095. eCollection 2022 Mar 18.
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).
血蓝蛋白 (KLH) 是一种糖基化的多亚基金属蛋白，能够引发强烈的非特异性免疫激活，从而诱导细胞和体液免疫反应。这种蛋白质的特殊免疫原性可以被利用来给小鼠接种自身抗原，否则这些自身抗原不会引起自身免疫反应。本方案描述了 KLH 与酰基辅酶 A 结合蛋白 (ACBP) 的共价偶联，以及用 ACBP-KLH 缀合物和一种有效的佐剂对小鼠进行自动免疫接种，并检测产生的抗 ACBP 自身抗体。有关此方案的使用和执行的完整详细信息，请参考 Bravo-San Pedro 等人 (2019c)。
