Measurement of thyroglobulin by liquid chromatography - Tandem mass spectrometry in serum and plasma in the presence of antithyroglobulin autoantibodies

Mark M. Kushnir, Alan L. Rockwood, William L. Roberts, Dev Abraham, Andrew N. Hoofnagle, A. Wayne Meikle

Research output: Contribution to journalArticlepeer-review

159 Scopus citations

Abstract

BACKGROUND: Measurement of serum thyroglobulin (Tg) is used to monitor patients after treatment for differentiated thyroid carcinoma (TC). Difficulty in using Tg as a biomarker of the recurrence of TC in many patients stems from the presence of endogenous anti-Tg autoantibodies (Tg-AAbs), which can interfere with immunoassays (IAs) and cause false-negative results. METHODS: We enriched Tg from serum samples using rabbit polyclonal anti-Tg antiserum and protein precipitation. Unrelated proteins were partially depleted in the process. Enriched proteins were then denatured, reduced, and digested with trypsin after the addition of a winged internal standard peptide. A Tg-specific tryptic peptide was purified by immunoaffinity extraction and analyzed by 2-dimensional LC-MS/MS. Instrument cycle time was 6.5 min per sample. RESULTS: The lower limit of quantification was 0.5 ng/mL (0.76 fmol/mL dimer). Total imprecision of triplicate measurements in serum samples over 5 days was
Original languageEnglish (US)
Pages (from-to)982-990
Number of pages9
JournalClinical Chemistry
Volume59
Issue number6
DOIs
StatePublished - Jun 1 2013

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

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