Capillary electrophoresis of methotrexate polyglutamates and its application in evaluation of γ-glutamyl hydrolase activity

Mark C. Waltham, Silan Lin, Wei Wei Li, Erdem Göker, Helena Gritsman, William P. Tong, Joseph R. Bertino

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A rapid and sensitive procedure for the separation of methotrexate (MTX) polyglutamates using capillary electrophoresis (CE) is described as it applies to the in vitro assay of the enzyme γ-glutamyl hydrolase (GGH, EC Distinct separation of MTX and polyglutamylated forms (up to glu4) is achieved within 10 min using a 75 μm I.D. capillary (50 cm, +25 kV), and enables quantitation of both reactant and enzyme products. As activity can be reliably determined using less than 5x105 eukaryotic cells, this new technique can be used to measure GGH in patient tumor samples and investigate the relationship between GGH levels and clinical MTX resistance.

Original languageEnglish
Pages (from-to)387-392
Number of pages6
JournalJournal of Chromatography B - Biomedical Applications
Issue number2
Publication statusPublished - 21 Feb 1997
Externally publishedYes


  • γ-Glutamyl hydrolase
  • 4-Amino-10-methylpteroylglutamate
  • Methotrexate polyglutamates

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