Rapid determination of multidrug resistance-associated protein in cancer cells by capillary electrophoresis immunoassay

Julius Mbuna, Takashi Kaneta, Totaro Imasaka

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

The adenosine triphosphate (ATP) binding-cassette (ABC) transporters are a superfamily of cellular proteins that have been partly implicated as a cause of multidrug resistance (MDR) in cancer cells. The ABC superfamily consists of P-glycoprotein, multidrug resistance-associated proteins (MRP) and breast cancer-related proteins, of which MRP is of particular interest because of its ability to efflux a broader range of substrates. Since MRP1 is the most prominent member of the MRP family, a simple technique is needed for its quantification. We developed a simple, fast (total analysis time of 3h) capillary electrophoresis immunoassay (CEIA) for the quantification of MRP1 in cancer cells. MRP1 antibody was labeled with fluorescein isothiocyanate. The labeled antibody was incubated with the cell lysate for a fixed interval (1h), after which the cell lysate mixture was directly injected into the capillary to separate the complex of MRP1 and its antibody from free antibody. The noncompetitive CEIA method had a limit of detection of 0.2nM and a good linear range (1.7-14.9×104 cells), and was fairly reproducible (RSD<10%). The results showed that two cell lines, A549 and RDES, expressed MRP1 in the absence of doxorubicin (DOX), with A549 registering a higher expression. Compared to DOX-free cancer cells, there was an acceleration of MRP1 expression during the 12h-exposure to DOX, after which the level of expression remained nearly constant as the intracellular accumulation of DOX decreased. The results obtained in this work indicate that the developed CEIA method is useful for relative quantification of MRPs in cancer cells.

Original languageEnglish
Pages (from-to)3923-3927
Number of pages5
JournalJournal of Chromatography A
Volume1218
Issue number25
DOIs
Publication statusPublished - Jun 24 2011

Keywords

  • Cancer cell
  • Capillary electrophoresis immunoassay
  • Doxorubicin
  • Laser-induced fluorescence
  • Multidrug resistance-associated protein

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

Fingerprint Dive into the research topics of 'Rapid determination of multidrug resistance-associated protein in cancer cells by capillary electrophoresis immunoassay'. Together they form a unique fingerprint.

  • Cite this