Related Experiment Video
Updated: Mar 10, 2026
Use of Capillary Electrophoresis Immunoassay to Search for Potential Biomarkers of Amyotrophic Lateral Sclerosis in Human Platelets
Published on: February 10, 2020
Quantification of ATP7B Protein in Dried Blood Spots by Peptide Immuno-SRM as a Potential Screen for Wilson's Disease
Sunhee Jung1, Jeffrey R Whiteaker2, Lei Zhao2
1Seattle Children's Hospital Research Institute , Seattle, Washington 98101, United States.
Insights
New methods can now detect ATP7B protein in dried blood spots, enabling newborn screening for Wilson Disease (WD). Early detection of this copper transport disorder can prevent severe neurological and liver damage.
Area of Science:
- Biochemistry
- Genetics
- Medical Diagnostics
Background:
- Wilson Disease (WD) is a genetic copper transport disorder linked to the ATP7B gene.
- Early intervention is crucial to prevent irreversible neurological and liver damage.
- Previous screening methods using ceruloplasmin (CP) have been insufficient for universal newborn screening (NBS).
Purpose of the Study:
- To evaluate ATP7B protein as a potential biomarker for Wilson Disease newborn screening.
- To develop and validate a method for quantifying ATP7B protein in dried blood spots (DBS).
Main Methods:
- Utilized an immuno-SRM platform to quantify ATP7B protein levels in DBS.
- Assessed assay precision, sensitivity, and protein stability in DBS.
Main Results:
- The immuno-SRM assay successfully quantified ATP7B in DBS at picomolar levels.
- The assay demonstrated high precision (<10% CV) and distinguished affected individuals from controls (p < 0.0001).
- ATP7B protein remained stable in DBS for one week at room temperature.
Conclusions:
- ATP7B protein quantification in DBS is a viable method for Wilson Disease newborn screening.
- This approach holds promise for early detection and prevention of severe WD complications.
- The platform may be adaptable for multiplexed screening of various congenital disorders using protein biomarkers.
Abstract:
Wilson's Disease (WD), a copper transport disorder caused by a genetic defect in the ATP7B gene, has been a long time strong candidate for newborn screening (NBS), since early interventions can give better results by preventing irreversible neurological disability or liver cirrhosis. Several previous pilot studies measuring ceruloplasmin (CP) in infants or children showed that this marker alone was insufficient to meet the universal screening for WD. WD results from mutations that cause absent or markedly diminished levels of ATP7B. Therefore, ATP7B could serve as a marker for the screening of WD, if the protein can be detected from dried blood spots (DBS). This study demonstrates that the immuno-SRM platform can quantify ATP7B in DBS in the picomolar range, and that the assay readily distinguishes affected cases from normal controls (p < 0.0001). The assay precision was <10% CV, and the protein was stable for a week in DBS at room temperature. These promising proof-of-concept data open up the possibility of screening WD in newborns and the potential for a multiplexed assay for screening a variety of congenital disorders using proteins as biomarkers in DBS.
