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.jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117 Biomed+Chromatogr
2018 ; 32
(8
): e4238
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Quantification of cystine in human renal proximal tubule cells using liquid
chromatography-tandem mass spectrometry
#MMPMID29517154
Jamalpoor A
; Sparidans RW
; Pou Casellas C
; Rood JJM
; Joshi M
; Masereeuw R
; Janssen MJ
Biomed Chromatogr
2018[Aug]; 32
(8
): e4238
PMID29517154
show ga
Nephropathic cystinosis is characterized by abnormal intralysosomal accumulation
of cystine throughout the body, causing irreversible damage to various organs,
particularly the kidneys. Cysteamine, the currently available treatment, can
reduce lysosomal cystine and postpone disease progression. However, cysteamine
poses serious side effects and does not address all of the symptoms of
cystinosis. To screen for new treatment options, a rapid and reliable
high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS)
method was developed to quantify cystine in conditionally immortalized human
proximal tubular epithelial cells (ciPTEC). The ciPTEC were treated with
N-ethylmaleimide, lysed and deproteinized with 15% (w/v) sulfosalicylic acid.
Subsequently, cystine was measured using deuterium-labeled cystine-D4, as the
internal standard. The assay developed demonstrated linearity to at least
20??mol/L with a good precision. Accuracies were between 97.3 and 102.9% for both
cell extracts and whole cell samples. Cystine was sufficiently stable under all
relevant analytical conditions. The assay was successfully applied to determine
cystine levels in both healthy and cystinotic ciPTEC. Control cells showed
clearly distinguishable cystine levels compared with cystinotic cells treated
with or without cysteamine. The method developed provides a fast and reliable
quantification of cystine, and is applicable to screen for potential drugs that
could reverse cystinotic symptoms in human kidney cells.
|Cell Line
[MESH]
|Chromatography, High Pressure Liquid/*methods
[MESH]