Selected Reaction Monitoring–Mass Spectrometric Immunoassay Responsive to Parathyroid Hormone and Related Variants

Lopez, Mary F.; Rezai, Taha; Sarracino, David; Prakash, Amol; Krastins, Bryan; Athanas, Michael; Singh, Ravinder J.; Barnidge, David R.; Oran, Paul E.; Borges, Chad R.; Nelson, Randall W.

doi:10.1373/clinchem.2009.137323

Cited by 117 publications

(119 citation statements)

References 27 publications

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“…All of these fragments, with the exception of PTH45-84, were also found in control samples. In 2010, Lopez et al [49] carried out similar experiments, again using immunoaffinity to enrich the PTH fragments. However, this time the immunoaffinity process was automated by having the antibody (polyclonal goat anti-human PTH39-84) immobilised on a micro-columns embedded into pipette tips [mass spectrometric immunoassay (MSIA)].…”

Section: Mass Spectrometrymentioning

confidence: 99%

“…Recently, two groups have published quantitative LC-MS/ MS methods for the analysis of PTH1-84 in clinical samples [44,49]. With these methods now available, it has been recognised that the development of a candidate reference method for PTH is now a possibility [16].…”

Section: Quantitative Lc-ms/ms Pth Assaysmentioning

confidence: 99%

“…With these methods now available, it has been recognised that the development of a candidate reference method for PTH is now a possibility [16]. Both methods utilise immunoaffinity extraction with a C-terminal anti-PTH capture antibody, either immobilised onto polystyrene beads [44] or onto customised MSIA pipette tips [49]. In both methods, following washing steps to remove nonspecific binding, the captured PTH1-84 and related fragments are digested using trypsin.…”

Section: Quantitative Lc-ms/ms Pth Assaysmentioning

confidence: 99%

“…Kumar et al [44] used isotopically labelled ( 15 N) PTH1-84 as the internal reference standard, whereas Lopez et al [49] used individual isotopically labelled tryptic peptides (one for each tryptic peptide monitored). Both groups used the quantification of the 1-13 amino acid tryptic peptide (SVSEIQLMHNLGK) as a surrogate for the measurement of PTH1-84.…”

Section: -84mentioning

confidence: 99%

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LC-MS candidate reference methods for the harmonisation of parathyroid hormone (PTH) measurement: a review of recent developments and future considerations

Couchman¹,

Taylor

Krastins

et al. 2014

Clinical Chemistry and Laboratory Medicine

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Abstract:The analysis of intact parathyroid hormone (PTH) (PTH1-84) is useful in the diagnosis of hyper-and hypocalcaemia, hyperparathyroidism, and in the prevention of bone mineral disorders in renal patients. The analysis is complicated by the presence of PTH fragments, which may accumulate in renal failure and cross-react in immunoassays, including the most recent third-generation immunoassays. Large variability exists between different commercially available assays. This article reviews the current literature on PTH testing, with emphasis on the use of mass spectrometry-based methods, and considers the important sources of variation which still need to be addressed prior to the development of much needed candidate reference methods for PTH analysis. Recently, mass spectrometric methods have been developed for the quantitation of PTH1-84 using surrogate tryptic peptides, but even these methods are subject to significant interferences due to the presence of newly observed modified PTH species, such as oxidised and phosphorylated PTH variants, which can accumulate in patient samples. Further work, including: 1) the use of high-resolution mass spectrometry; and 2) the analysis of PTH without prior protease digestion, is required before these approaches can be considered as reference methods against which other methods should be harmonised.

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Section: Mass Spectrometrymentioning

confidence: 99%

Section: Quantitative Lc-ms/ms Pth Assaysmentioning

confidence: 99%

Section: Quantitative Lc-ms/ms Pth Assaysmentioning

confidence: 99%

Section: -84mentioning

confidence: 99%

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LC-MS candidate reference methods for the harmonisation of parathyroid hormone (PTH) measurement: a review of recent developments and future considerations

Couchman¹,

Taylor

Krastins

et al. 2014

Clinical Chemistry and Laboratory Medicine

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“…Even with PSA enrichment by antibody affinity chromatography, the sensitivity difference is 1000-fold [5]. Some newer affinity purification strategies may be promising, if coupled to SRM, but they are currently more complex, time-consuming and lower throughput than ELISA [6].…”

Section: Biomarker Validation Is Still the Bottleneck In Biomarker Rementioning

confidence: 99%

Biomarker validation is still the bottleneck in biomarker research

Diamandis

2012

J Intern Med

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Absolute Quantification of Proteins by Mass Spectrometry

Shuford

Muddiman

2000

Encyclopedia of Analytical Chemistry

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Mass spectrometry (MS) provides unambiguous molecular specificity relative to all other analytical and immunological techniques and, consequently, has great potential to provide unambiguous estimates of absolute protein quantities within complex biological systems. Here, we provide a critical review and discussion of the predominant MS ‐based technology used for absolute protein quantification, referred to as protein cleavage‐isotope dilution mass spectrometry ( PC‐IDMS ). This technique requires proteolytic conversion of the analyte protein into its quantitative surrogate peptide, which is subsequently quantified using a stable isotope‐labeled (SIL) peptide analog as an internal standard (IS). All phases of the PC‐IDMS workflow, from the SIL peptide production platforms to data analysis, are described and characterized in detail. An emphasis is placed on the practical quantitative aspects for each step of the workflow and their implication on quantitative accuracy.

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Selected Reaction Monitoring–Mass Spectrometric Immunoassay Responsive to Parathyroid Hormone and Related Variants

Cited by 117 publications

References 27 publications

LC-MS candidate reference methods for the harmonisation of parathyroid hormone (PTH) measurement: a review of recent developments and future considerations

LC-MS candidate reference methods for the harmonisation of parathyroid hormone (PTH) measurement: a review of recent developments and future considerations

Biomarker validation is still the bottleneck in biomarker research

Absolute Quantification of Proteins by Mass Spectrometry

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