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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Lopez, Mary F. Rezai, Taha Sarracino, David A. Prakash, Amol Krastins, Bryan Athanas, Michael Singh, Ravinder J. Barnidge, David R. Oran, Paul Borges, Chad Nelson, Randall W. |
| Description | Country affiliation: United States Author Affiliation: Lopez MF ( ThermoFisher Scientific BRIMS, Cambridge, MA, USA.) |
| Abstract | BACKGROUND: Parathyroid hormone (PTH) assays able to distinguish between full-length PTH (PTH1-84) and N-terminally truncated PTH (PTH7-84) are of increasing significance in the accurate diagnosis of endocrine and osteological diseases. We describe the discovery of new N-terminal and C-terminal PTH variants and the development of selected reaction monitoring (SRM)-based immunoassays specifically designed for the detection of full-length PTH [amino acid (aa)1-84] and 2 N-terminal variants, aa7-84 and aa34-84. METHODS: Preparation of mass spectrometric immunoassay pipettor tips and MALDI-TOF mass spectrometric analysis were carried out as previously described. We used novel software to develop SRM assays on a triple-quadrupole mass spectrometer. Heavy isotope-labeled versions of target peptides were used as internal standards. RESULTS: Top-down analysis of samples from healthy individuals and renal failure patients revealed numerous PTH variants, including previously unidentified aa28-84, aa48-84, aa34-77, aa37-77, and aa38-77. Quantitative SRM assays were developed for PTH1-84, PTH7-84, and variant aa34-84. Peptides exhibited linear responses (R(2) = 0.90-0.99) relative to recombinant human PTH concentration limits of detection for intact PTH of 8 ng/L and limits of quantification of 16-31 ng/L depending on the peptide. Standard error of analysis for all triplicate measurements was 3%-12% for all peptides, with <5% chromatographic drift between replicates. The CVs of integrated areas under the curve for 54 separate measurements of heavy peptides were 5%-9%. CONCLUSIONS: Mass spectrometric immunoassays identified new clinical variants of PTH and provided a quantitative assay for these and previously identified forms of PTH. |
| File Format | HTM / HTML |
| ISSN | 00099147 |
| e-ISSN | 15308561 |
| Journal | Clinical Chemistry |
| Issue Number | 2 |
| Volume Number | 56 |
| Language | English |
| Publisher | American Association for Clinical Chemistry |
| Publisher Date | 2010-02-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Research Support, N.i.h., Extramural Immunoassay Amino Acid Sequence Peptide Fragments Area Under Curve Molecular Sequence Data Discipline Laboratory Medicine Parathyroid Hormone Blood Spectrometry, Mass, Matrix-assisted Laser Desorption-ionization Limit Of Detection Discipline Clinical Chemistry Kidney Failure, Chronic Diagnosis |
| Content Type | Text |
| Resource Type | Article |
| Subject | Biochemistry (medical) Clinical Biochemistry |
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