A Bead-Based Nonradioactive Immunoassay for Autoantibody Testing in a Mouse Model of Myasthenia Gravis

IF 3 Q3 IMMUNOLOGY

Antibodies Pub Date : 2024-07-01 DOI:10.3390/antib13030053

Afrin Bahauddin, K. Curtis, J. Guptarak, R. Huda

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引用次数: 0

Abstract

Serological testing for anti-acetylcholine receptor (AChR) autoantibodies is not only crucial for the diagnosing, disease monitoring, and treatment management of patients with myasthenia gravis (MG) but also for preclinical studies utilizing MG disease models. However, there are no specific guidelines on which methods to use in clinical diagnostic or research laboratories to detect or quantify any MG-specific autoantibodies. Conventional autoantibody assays, particularly those for anti-AChR antibodies, are varied and mostly laboratory-specific. Here, we report our new nonradioactive immunoprecipitation–immunoblotting method for assessing autoantibodies (anti-AChR antibodies) in a mouse model of MG. This simple, efficient, reproducible, and cost-effective assay appears superior to the enzyme-linked immunosorbent assay but comparable to the radioimmunoprecipitation or cell-based assay in specificity and sensitivity. Thus, the newly developed assay can serve as a valuable alternative to classical assays and is suitable for routine testing of AChR-specific autoantibodies in preclinical studies. The further optimization of our assay may facilitate its application in the diagnosis and therapeutic management of patients with MG.

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基于微珠的非放射性免疫测定法，用于肌无力小鼠模型的自身抗体检测

抗乙酰胆碱受体（AChR）自身抗体的血清学检测不仅对重症肌无力（MG）患者的诊断、疾病监测和治疗管理至关重要，而且对利用MG疾病模型进行临床前研究也很重要。然而，对于临床诊断或研究实验室使用哪种方法检测或量化任何 MG 特异性自身抗体，目前还没有具体的指导原则。传统的自身抗体检测方法，尤其是抗 AChR 抗体的检测方法多种多样，而且大多具有实验室特异性。在此，我们报告了一种新的非放射性免疫沉淀-免疫印迹法，用于评估小鼠 MG 模型中的自身抗体（抗 AChR 抗体）。这种简单、高效、可重复、经济的检测方法优于酶联免疫吸附检测法，但在特异性和灵敏度方面与放射免疫沉淀或细胞检测法不相上下。因此，新开发的检测方法可作为经典检测方法的重要替代方法，适用于临床前研究中 AChR 特异性自身抗体的常规检测。进一步优化我们的检测方法可能有助于将其应用于 MG 患者的诊断和治疗管理。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Antibodies IMMUNOLOGY-

CiteScore

7.10

自引率

6.40%

发文量

审稿时长

11 weeks

期刊介绍： Antibodies (ISSN 2073-4468), an international, peer-reviewed open access journal which provides an advanced forum for studies related to antibodies and antigens. It publishes reviews, research articles, communications and short notes. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. Full experimental and/or methodical details must be provided. Electronic files or software regarding the full details of the calculation and experimental procedure - if unable to be published in a normal way - can be deposited as supplementary material. This journal covers all topics related to antibodies and antigens, topics of interest include (but are not limited to): antibody-producing cells (including B cells), antibody structure and function, antibody-antigen interactions, Fc receptors, antibody manufacturing antibody engineering, antibody therapy, immunoassays, antibody diagnosis, tissue antigens, exogenous antigens, endogenous antigens, autoantigens, monoclonal antibodies, natural antibodies, humoral immune responses, immunoregulatory molecules.