Summary
An isotachophoretic method using polyacrylamide gel (PAG-ITP) in a simple disc electrophoretic equipment with plastic tubes containing the gels, was elaborated and especially designed for studying the gammaglobulins in CSF and serum from control subjects and patients with neurological disorders, especially known or probable MS. The device and the ITP system used, including leading and terminating electrolytes and spacer substances, dividing the gammalgobulins in a reproducible way, are described. No cooling of the gel tubes was needed. The sample volumes varied between 5–500 μl, and the separation time was 1.5–3.0 h. CSF from patients with verified or probable MS revealed characteristic, increased low-mobility gammaglobulin fractions.
Using other ITP systems, such as other spacer compositions, the anodic proteins can also be studied in more detail. PAG-ITP in gel tubes is a simple and inexpensive technique which can be used for both analytical and preparative procedures for biological material such as CSF, serum and extractions from nervous tissues.
Zusammenfassung
Es wird eine isotachophoretische Methode beschrieben, welche Polyacrylamidgel (PAG-ITP) in einem einfachen selbstgebauten Scheibenelektrophoresegerät mit Kunststoffröhrchen benutzt. Die Methode ist besonders geeignet zum Studium von Gammaglobulin im Liquor und Serum, sowohl von Normalfällen wie auch bei Patienten mit neurologischen Krankheiten, im besonderen mit Multipler Sklerose. Es wird sowohl das Gerät selber beschrieben, wie auch das isotachophoretische (ITP) System, inklusive die Leiter und die Endelektrolyte sowie die Trennstoffe. Das ITP-System trennt die Gammaglobuline in reproduzierbarer Weise. Eine Kühlung der Gelröhrchen ist nicht notwendig. Das Volumen der untersuchten Proben schwankte zwischen 5 und 100 μl, die Trennzeit zwischen 1,5 und 3 Std. Im Liquor von gesicherten oder möglichen MS-Fällen zeigten sich charakteristische, langsam wandernde Gammaglobulinfraktionen. Mittels anderer ITP-Systeme oder anderer Trennstoffmischungen können auch die anodischen Proteine im Detail untersucht werden. PAG-ITP in Gelröhrchen stellt eine einfache und billige Untersuchungstechnik dar. Sie kann sowohl analytisch wie präparativ verwendet werden, für biologische Proben wie für Liquor, Serum und Extrakte aus Nervengewebe.
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Kjellin, K.G., Hallander, L. Isotachophoresis of CSF proteins in gel tubes especially gammaglobulins. J Neurol 221, 225–233 (1979). https://doi.org/10.1007/BF00314639
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DOI: https://doi.org/10.1007/BF00314639