Kleine Tilmann O, Damm Thomas
Department of Clinical Chemistry and Molecular Diagnostics, Clinicum of the University Marburg, D-35033 Marburg, Germany.
Brain Res Bull. 2003 Aug 15;61(3):309-20. doi: 10.1016/s0361-9230(03)00094-7.
An optimized automated IEF procedure in polyacrylamide micro gels and immunofixation with 10 monospecific antibodies against some fragments of the IgG molecule and against the whole IgG isotypes IgG(1), IgG(2), IgG(3), IgG(4), respectively, detected oligoclonal bands (OBs) within acid, neutral, and alkaline ranges of the gels. Accuracy and reliability of the OB assay for detection of an intrathecal IgG synthesis proved to be higher with immunofixation than with silver staining of bands precipitated by trichloroacetic acid. CSF OBs were specified as lambda or kappa IgG subfractions, respectively, duplex IgGs precipitated with anti-lambda, anti-kappa, and anti-Fc antibodies at the same pI. Most of OBs were classified as belonging to either IgG(1), IgG(2), IgG(3), or IgG(4) isotypes in CSF. The IEF procedure additionally allowed the discrimination of both free light chains and possible "free" heavy gamma chain fragments in CSF, when immunofixation was done with monospecific antibodies against both light chains and gamma chain fragments (e.g. anti-F(ab')(2), anti-Fd, anti-Fc, anti-C(H)2). The results pointed out a distinct heterogeneity of the IgG immune response in human CSF. The IEF procedure with extended immunofixation is recommended for completion of the basic laboratory procedures used in neuroimmunology in order to discriminate inflammatory processes in human CNS.
一种优化的聚丙烯酰胺微凝胶自动等电聚焦程序，并用针对IgG分子某些片段以及分别针对整个IgG同种型IgG(1)、IgG(2)、IgG(3)、IgG(4)的10种单特异性抗体进行免疫固定，在凝胶的酸性、中性和碱性范围内检测到寡克隆带(OBs)。结果证明，免疫固定法检测鞘内IgG合成的OB测定法的准确性和可靠性高于三氯乙酸沉淀带的银染法。CSF OBs分别被指定为λ或κIgG亚组分，即在相同pI下用抗λ、抗κ和抗Fc抗体沉淀的双链IgG。CSF中的大多数OBs被分类为属于IgG(1)、IgG(2)、IgG(3)或IgG(4)同种型。当用针对轻链和γ链片段的单特异性抗体（例如抗F(ab')(2)、抗Fd、抗Fc、抗C(H)2）进行免疫固定时，等电聚焦程序还可以区分CSF中的游离轻链和可能的“游离”重γ链片段。结果指出了人CSF中IgG免疫反应的明显异质性。建议采用延长免疫固定的等电聚焦程序来完成神经免疫学中用于区分人类中枢神经系统炎症过程的基本实验室程序。
