Multicenter evaluation of a novel nanoparticle immunoassay for 5-fluorouracil on the olympus AU400 analyzer

Jan H. Beumer, M. Boisdron-Celle, William Clarke, Jodi B. Courtney, Merrill J. Egorin, Erick Gamelin, Rebecca L. Harney, Catherine Hammett-Stabler, Sandy Lepp, Yunying Li, Gregory D. Lundell, Gwen McMillin, Gerard Milano, Salvatore J. Salamone

Research output: Contribution to journalArticlepeer-review

36 Scopus citations

Abstract

Background: 5-Fluorouracil (5-FU) is the most widely used chemotherapy drug, primarily against gastrointestinal, head and neck, and breast cancers. 5-FU has large pharmacokinetic variability resulting in unexpected toxicity or ineffective treatment. Therapeutic drug management of 5-FU minimizes toxicity and improves outcome. A nanoparticle-based immunoassay was developed to provide oncologists with a rapid, cost-effective tool for determining 5-FU plasma concentrations. METHODS: Monoclonal antibodies, bound to nanoparticles, were used to develop an immunoassay for the Olympus AU400. Assay precision, linearity, calibration stability, and limit of detection were run at multiple centers; interference, cross-reactivity, lower limit of quantitation and recovery at 1 center. Clinical samples collected from 4 cancer centers were analyzed for 5-FU concentrations by liquid chromatography-tandem mass spectrometry and compared with the immunoassay results. RESULTS: With calibrators from 0 to 1800 ng/mL 5-FU and autodilution, concentrations up to 9000 ng/mL could be determined. Time to first result was 10 minutes, and 400 samples per hour could be quantitated from a standard curve stored for >30 days. Imprecision across all laboratories was <5%, and the assay was linear upon dilution over the entire range. Cross-reactivities for dihydro-5-FU, uracil, capecitabine, and tegafur were <1%, 9.9%, 0.05%, and 0.23%, respectively. The limit of detection was 52 ng/mL with a lower limit of quantitation of 86 ng/mL. Assay results of clinical samples (93-1774 ng/mL) correlated with liquid chromatography-tandem mass spectrometry results: (R = 0.9860, slope 1.035, intercept 10.87 ng/mL). CONCLUSIONS: This novel immunoassay is suitable for quantitating 5-FU plasma concentrations with advantages of speed, small sample size, minimal sample pretreatment, and application on automated instrumentation. These advantages enable efficient therapeutic drug management of 5-FU in clinical practice.

Original languageEnglish (US)
Pages (from-to)688-694
Number of pages7
JournalTherapeutic drug monitoring
Volume31
Issue number6
DOIs
StatePublished - Dec 2009

Keywords

  • 5-fluorouracil
  • Chemotherapy
  • Drug management
  • Immunoassay

ASJC Scopus subject areas

  • Pharmacology
  • Pharmacology (medical)

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