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 <10%. Comparison with a commercial IA using serum samples free of Tg-AAb (n = 73) showed Deming regression, IA = 1.00 * LC-MS/MS - 2.35, r = 0.982, standard error of the estimate (S(y|x)) = 9.52. In a set of Tg-AAb-positive samples that tested negative for Tg using IA (n = 71), concentrations determined by LC-MS/MS were ≥0.5 ng/mL in 23% of samples (median 1.2, range 0.7-11 ng/mL). CONCLUSIONS:The introduced method has acceptable performance characteristics for use in clinical diagnostic applications. The most substantial disagreement between methods was observed in Tg-AAb-positive samples with concentrations <2 ng/mL (determined with LC-MS/MS). The affinity-assisted enrichment strategy used for Tg in this method should be applicable to other biomarkers that have endogenous autoantibodies.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Kushnir MM,Rockwood AL,Roberts WL,Abraham D,Hoofnagle AN,Meikle AWdoi
10.1373/clinchem.2012.195594subject
Has Abstractpub_date
2013-06-01 00:00:00pages
982-90issue
6eissn
0009-9147issn
1530-8561pii
clinchem.2012.195594journal_volume
59pub_type
杂志文章abstract：:The concentration of the conjugated bile acid, cholylglycine, in serum is a sensitive and specific indicator of hepatic function. We describe a convenient, specific, and precise radioimmunoassay for cholylglycine, in which 125I-labeled cholylglycyltyrosine is used as tracer. In addition, a blocking agent in the buffer...
journal_title：Clinical chemistry
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abstract：:We have developed and evaluated a new procedure for detecting trends in quality-control measurements and applied it to laboratory data. The method requires the use of sequential or "moving" slope estimates to identify trends. Formulae are derived to estimate the regression error for the moving slope directly from the ...
journal_title：Clinical chemistry
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journal_title：Clinical chemistry
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journal_title：Clinical chemistry
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journal_title：Clinical chemistry
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doi：
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更新日期：1982-05-01 00:00:00
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doi：
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doi：
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