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改良放射免疫测定法与 ELISA 定量检测重症肌无力小鼠模型中的抗乙酰胆碱受体抗体。

Modified radioimmunoassay versus ELISA to quantify anti-acetylcholine receptor antibodies in a mouse model of myasthenia gravis.

机构信息

Immunology Department, Hospital de la Santa Creu i Sant Pau, Biomedical Research Institute Sant Pau (IIB Sant Pau), Barcelona, Spain.

AHEAD Therapeutics S.L. Barcelona, Spain.

出版信息

J Immunol Methods. 2024 Nov;534:113748. doi: 10.1016/j.jim.2024.113748. Epub 2024 Sep 4.

DOI:10.1016/j.jim.2024.113748
PMID:39241980
Abstract

In mouse models of myasthenia gravis (MG), anti-acetylcholine receptor (AChR) antibodies can be quantified to monitor disease progression and treatment response. In mice, enzyme-linked immunosorbent assay (ELISA) is the gold standard to quantify these antibodies. However, this method requires antigen purification, which is both time-consuming and expensive. In humans, radioimmunoassay (RIA)-which is more sensitive than ELISA-is commonly used to quantify AChR antibodies. At present, however, no commercial RIA kits are available to quantify these antibodies in mice. The aim of this study was to compare a modified commercial human RIA kit to two ELISA methods to detect AChR antibodies in an experimental autoimmune mouse model of MG (EAMG). C57BL/6 J mice were immunized with purified AChR from Tetronarce californica (T-AChR). Serum samples were analyzed by RIA and two ELISAs (T-AChR and purified mouse AChR peptide [m-AChR]). The modified RIA showed excellent sensitivity (84.1 %) and specificity (100 %) for the detection of AChR antibodies. RIA showed a good agreement with T-AChR ELISA (κ = 0.69) but only moderate agreement with m-AChR ELISA (κ = 0.49). These results demonstrate the feasibility of modifying a commercially-available RIA kit to quantify AChR antibodies in EAMG. The advantage of this technique is that it eliminates the need to develop the entire methodology in-house and reduces inter and intra-laboratory variability.

摘要

在重症肌无力(MG)的小鼠模型中,可以定量检测抗乙酰胆碱受体(AChR)抗体来监测疾病进展和治疗反应。在小鼠中,酶联免疫吸附测定(ELISA)是定量这些抗体的金标准。然而,这种方法需要抗原纯化,既费时又昂贵。在人类中,放射免疫分析(RIA)——比 ELISA 更敏感——常用于定量 AChR 抗体。然而,目前没有商业 RIA 试剂盒可用于定量小鼠中的这些抗体。本研究旨在比较改良的商业人类 RIA 试剂盒与两种 ELISA 方法,以检测实验性自身免疫性 MG (EAMG)小鼠模型中的 AChR 抗体。C57BL/6J 小鼠用纯化的来自 Tetronarce californica 的 AChR(T-AChR)免疫。通过 RIA 和两种 ELISA(T-AChR 和纯化的小鼠 AChR 肽 [m-AChR])分析血清样本。改良的 RIA 对 AChR 抗体的检测具有出色的灵敏度(84.1%)和特异性(100%)。RIA 与 T-AChR ELISA 具有良好的一致性(κ=0.69),但与 m-AChR ELISA 仅有中等一致性(κ=0.49)。这些结果表明,改良商业 RIA 试剂盒来定量 EAMG 中的 AChR 抗体是可行的。该技术的优势在于它消除了在内部开发整个方法的需要,并减少了实验室间和实验室内部的变异性。

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