A distributable LC-MS/MS method for the measurement of serum thyroglobulin
•Sensitive serum thyroglobulin measurement by LC-MS/MS.•Detailed standard operating procedure using a commercially available antibody.•Dimethylsulfoxide-enhanced ionization of a proteotypic thyroglobulin peptide.•Beads for peptide immunoaffinity purification can be reused at least 5 times.•Distribut...
Saved in:
Published in | Journal of mass spectrometry and advances in the clinical lab Vol. 26; pp. 28 - 33 |
---|---|
Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
Netherlands
Elsevier B.V
01.11.2022
Elsevier |
Subjects | |
Online Access | Get full text |
Cover
Loading…
Summary: | •Sensitive serum thyroglobulin measurement by LC-MS/MS.•Detailed standard operating procedure using a commercially available antibody.•Dimethylsulfoxide-enhanced ionization of a proteotypic thyroglobulin peptide.•Beads for peptide immunoaffinity purification can be reused at least 5 times.•Distributable standard reference material (Husky Ref) to facilitate assay harmonization.
Despite its clear advantages over immunoassay-based testing, the measurement of serum thyroglobulin by mass spectrometry remains limited to a handful of institutions. Slow adoption by clinical laboratories could reflect limited accessibility to existing methods that have sensitivity comparable to modern immunoassays, as well as a lack of tools for calibration and assay harmonization.
We developed and validated a liquid chromatography-tandem mass spectrometry-based assay for the quantification of serum thyroglobulin. The protocol combined peptide immunoaffinity purification using a commercially available, well-characterized monoclonal antibody and mobile phase modification with dimethylsulfoxide (DMSO) for enhanced sensitivity. To facilitate harmonization with other laboratories, we developed a novel, serum-based 5-point distributable reference material (Husky Ref).
The assay demonstrated a lower limit of quantification of 0.15 ng/mL (<20 %CV). Mobile phase DMSO increased signal intensity of the target peptide at least 3-fold, improving quantification at low concentrations. Calibration traceable to Husky Ref enabled harmonization between laboratories in an interlaboratory study.
Sensitive mass spectrometry-based thyroglobulin measurement can be achieved using a monoclonal antibody during peptide immunoaffinity purification and the addition of mobile phase DMSO. Laboratories interested in deploying this assay can utilize the provided standard operating procedure and freely-available Husky Ref reference material. |
---|---|
Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Present addresses: Department of Pathology and Laboratory Medicine, Vancouver Coastal Health; Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada (JS); Labcorp, Burlington, NC (BYO); Seagen, Inc., Bothel, WA (CMH), USA. |
ISSN: | 2667-145X 2667-1468 2667-145X |
DOI: | 10.1016/j.jmsacl.2022.09.005 |