Abstract：
BACKGROUND:Cystinosis is a rare autosomal recessive disorder characterized by an accumulation of intralysosomal cystine due to a defect in cystine transport across the lysosomal membrane. This disorder can be treated specifically using high doses of cysteamine. Accurate measurement of intracellular cystine content is necessary for the diagnosis and monitoring of treatment with cysteamine. Here we describe a new method to measure intracellular cystine. It relies on a liquid chromatography-tandem mass spectrometry assay. We compare this novel method with the cystine-binding protein assay. METHOD:Cells were isolated and lysed in the presence of N-ethylmaleimide to avoid interference from cysteine. After deproteinization, addition of stable isotope d6 cystine and butylation, cystine was measured using an API 3000 MSMS. RESULTS:The cystine assay was linear to at least 50 micromol/L. Within-run and between-run coefficients of variation were 2.9% and 5.7% respectively. CONCLUSION:It is possible to measure very low concentrations of intracellular cystine with liquid chromatography-tandem mass spectrometry. The results obtained with this novel method correlate very well with those obtained using the cystine-binding protein assay.
journal_name
Clin Biochemjournal_title
Clinical biochemistryauthors
Chabli A,Aupetit J,Raehm M,Ricquier D,Chadefaux-Vekemans Bdoi
10.1016/j.clinbiochem.2007.02.005subject
Has Abstractpub_date
2007-06-01 00:00:00pages
692-8issue
9-10eissn
0009-9120issn
1873-2933pii
S0009-9120(07)00098-7journal_volume
40pub_type
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