Cowdrey G, Gould B, Rees J, Firth G
Department of Biochemistry, Hurstwood Park Neurological Centre, Haywards Heath, Sussex, UK.
Electrophoresis. 1990 Oct;11(10):813-8. doi: 10.1002/elps.1150111007.
A method is described for the separation and detection of highly alkaline IgG bands in unconcentrated cerebrospinal fluid (CSF). These bands are frequently found in the cerebrospinal fluid of patients with inflammatory diseases of the central nervous system, particularly in the case of multiple sclerosis, and their detection is an important aid in clinical diagnosis. An isoelectric focusing technique using an immobilised pH gradient in polyacrylamide gel has been developed over the pH range 7-10, producing a linear and stable pH gradient with excellent resolution. After electrofocusing, the protein patterns were blotted onto polyvinylidene difluoride membranes and visualised using anti-human IgG followed by an enzyme-labelled second antibody. Blotting could be carried out by capillary diffusion for up to 16 h duration without any loss in resolution. Using this method, highly alkaline intrathecal IgG bands were found in the cerebrospinal fluid of all of the 14 multiple sclerosis patients. There were also 2 patients with alkaline IgG bands in their cerebrospinal fluid who were not diagnosed as multiple sclerosis. By contrast, no alkaline IgG bands with an isoelectric point (pI) greater than 8.6 were found in any of the serum samples studied (n = 50) from patients with various neurological disorders including multiple sclerosis.
本文描述了一种用于分离和检测未浓缩脑脊液（CSF）中高碱性IgG条带的方法。这些条带常见于中枢神经系统炎症性疾病患者的脑脊液中，尤其是在多发性硬化症患者中，其检测对临床诊断有重要帮助。已开发出一种在聚丙烯酰胺凝胶中使用固定化pH梯度的等电聚焦技术，pH范围为7 - 10，可产生线性且稳定的pH梯度，分辨率极佳。电聚焦后，将蛋白质图谱印迹到聚偏二氟乙烯膜上，先用抗人IgG进行检测，再用酶标记的二抗进行显色。印迹可通过毛细管扩散进行长达16小时，分辨率无任何损失。使用该方法，在14例多发性硬化症患者的脑脊液中均发现了高碱性鞘内IgG条带。还有2例脑脊液中有碱性IgG条带的患者未被诊断为多发性硬化症。相比之下，在包括多发性硬化症在内的各种神经系统疾病患者的任何血清样本（n = 50）中均未发现等电点（pI）大于8.6的碱性IgG条带。
