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

doi:10.1016/S0378-4347(96)00350-7

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

Cancer Systems Imaging

Research output: Contribution to journal › Article › peer-review

13 Scopus citations

Abstract

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 3.4.22.12). 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 language	English (US)
Pages (from-to)	387-392
Number of pages	6
Journal	Journal of Chromatography B: Biomedical Applications
Volume	689
Issue number	2
DOIs	https://doi.org/10.1016/S0378-4347(96)00350-7
State	Published - Feb 21 1997

Keywords

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

ASJC Scopus subject areas

General Chemistry

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10.1016/S0378-4347(96)00350-7

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@article{e8d2c91bb620465fbdc8992c2d8f7555,

title = "Capillary electrophoresis of methotrexate polyglutamates and its application in evaluation of γ-glutamyl hydrolase activity",

abstract = "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 3.4.22.12). 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.",

keywords = "4-Amino-10-methylpteroylglutamate, Methotrexate polyglutamates, γ-Glutamyl hydrolase",

author = "Waltham, {Mark C.} and Silan Lin and Li, {Wei Wei} and Erdem G{\"o}ker and Helena Gritsman and Tong, {William P.} and Bertino, {Joseph R.}",

note = "Funding Information: We are currently investigating blast samples obtained from patients pre-and post-chemotherapy for regimens which have included antifolates. Together with determined activity measurements for folylpolyglutamate synthetase in these extracts, these analyses may shed light on the observations that variable antifolate polyglutamylation potential exists for different tumor varieties. In addition, and in This work was supported by grants from the United States Public Health Service (CA08010) and the American Cancer Society (ACS DHP-18-J).",

year = "1997",

month = feb,

day = "21",

doi = "10.1016/S0378-4347(96)00350-7",

language = "English (US)",

volume = "689",

pages = "387--392",

journal = "Journal of Chromatography B: Biomedical Applications",

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T1 - Capillary electrophoresis of methotrexate polyglutamates and its application in evaluation of γ-glutamyl hydrolase activity

AU - Waltham, Mark C.

AU - Lin, Silan

AU - Li, Wei Wei

AU - Göker, Erdem

AU - Gritsman, Helena

AU - Tong, William P.

AU - Bertino, Joseph R.

N1 - Funding Information: We are currently investigating blast samples obtained from patients pre-and post-chemotherapy for regimens which have included antifolates. Together with determined activity measurements for folylpolyglutamate synthetase in these extracts, these analyses may shed light on the observations that variable antifolate polyglutamylation potential exists for different tumor varieties. In addition, and in This work was supported by grants from the United States Public Health Service (CA08010) and the American Cancer Society (ACS DHP-18-J).

PY - 1997/2/21

Y1 - 1997/2/21

N2 - 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 3.4.22.12). 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.

AB - 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 3.4.22.12). 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.

KW - 4-Amino-10-methylpteroylglutamate

KW - Methotrexate polyglutamates

KW - γ-Glutamyl hydrolase

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

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