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一种基于酶联免疫吸附测定法对高粘性血清样本进行抗糖脂抗体评估的新方法。

A new approach to ELISA-based anti-glycolipid antibody evaluation of highly adhesive serum samples.

作者信息

Usuki Seigo, O'Brien Dawn, Rivner Michael H, Yu Robert K

机构信息

Department of Neuroscience and Regenerative Medicine, Medical College of Georgia, Georgia Regents University, Augusta, GA 30912, USA.

Department of Neurology, Medical College of Georgia, Georgia Regents University, Augusta, GA 30912, USA.

出版信息

J Immunol Methods. 2014 Jun;408:52-63. doi: 10.1016/j.jim.2014.05.005. Epub 2014 May 23.

Abstract

The enzyme-linked immunosorbent assay (ELISA) is a standard immunoassay used in measuring antibody reactivity (expressed as titers) for glycosphingolipids (GSLs) such as gangliosides and sulfoglycolipids in the sera of patients with Guillain-Barré syndrome (GBS), variants of GBS, and chronic inflammatory demyelinating polyneuropathy (CIDP). In the present study, anti-GSL antibodies were evaluated using a new formula of affinity parametric complex (APC), calculated from limiting-dilution serum assay data, followed by affinity parametric complex criterion (APCC). Using assay results based on APCC, we analyzed serum samples categorized into acute inflammatory demyelinating polyneuropathy (AIDP), acute motor-sensory axonal neuropathy (AMSAN), CIDP, CIDP with myasthenia gravis (MG), and amyotrophic lateral sclerosis (ALS). We were able to determine the affinity strength of antibodies otherwise hidden in the non-specific background activity in highly adhesive serum samples. The thin-layer chromatography (TLC)-immuno-overlay method assured us that this new method is an accurate and reliable way for evaluating anti-GSL antibodies using ELISA serum sample data.

摘要

酶联免疫吸附测定(ELISA)是一种标准免疫测定法,用于测量吉兰-巴雷综合征(GBS)、GBS变异型和慢性炎症性脱髓鞘性多发性神经病(CIDP)患者血清中神经节苷脂和硫苷脂等糖鞘脂(GSL)的抗体反应性(以滴度表示)。在本研究中,使用从有限稀释血清测定数据计算得出的亲和参数复合物(APC)新公式,随后采用亲和参数复合物标准(APCC)来评估抗GSL抗体。利用基于APCC的测定结果,我们分析了分类为急性炎症性脱髓鞘性多发性神经病(AIDP)、急性运动感觉轴索性神经病(AMSAN)、CIDP、合并重症肌无力(MG)的CIDP以及肌萎缩侧索硬化症(ALS)的血清样本。我们能够确定在高粘性血清样本中非特异性背景活性中隐藏的抗体的亲和强度。薄层色谱(TLC)免疫覆盖法向我们保证,这种新方法是一种利用ELISA血清样本数据评估抗GSL抗体的准确可靠方法。

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