Detection of α-Galactosidase A Reaction in Samples Extracted from Dried Blood Spots Using Ion-Sensitive Field Effect...

Area of Science:

• Biomedical Engineering
• Analytical Chemistry
• Biochemistry

Background:

• Fabry disease is a lysosomal storage disorder linked to reduced alpha-galactosidase A activity.
• Newborn screening for Fabry disease is feasible due to available enzyme replacement therapies.
• Current diagnostic methods may require improvement for widespread newborn screening.

Purpose of the Study:

• To develop and validate an electrochemical method for detecting alpha-galactosidase A activity in dried blood spots.
• To utilize CMOS-compatible ion-sensitive field-effect transistors (ISFETs) for label-free enzyme detection.
• To assess the potential of this method for enhancing newborn screening of Fabry disease.

Main Methods:

• An electrochemical detection method employing CMOS-compatible ISFETs with hafnium oxide surfaces was developed.
• Buffer composition was optimized for both enzyme activity and ISFET performance.
• Label-free detection of alpha-galactosidase A was achieved using its natural substrate, melibiose, in dried blood spot extracts.

Main Results:

• The ISFETs successfully detected the enzymatic reaction catalyzed by alpha-galactosidase A.
• Optimized buffer conditions supported both enzyme function and sensor performance.
• Packaged ISFET sensors effectively discriminated between normal and inhibited enzyme activity in dried blood samples.

Conclusions:

• The developed ISFET-based electrochemical method provides a promising approach for Fabry disease newborn screening.
• Label-free detection using natural substrates offers an advantage over traditional methods.
• This technology has the potential to increase the accessibility and distribution of newborn screening for Fabry disease.