A Chemiluminescent Protein Microarray Method for Determining the Seroglycoid Fucosylation Index

Area of Science:

• Biochemistry
• Oncology
• Medical Diagnostics

Background:

• Hepatocellular carcinoma (HCC) screening relies on biomarkers like Lens culinaris agglutinin-reactive alpha-fetoprotein (AFP-L3).
• Accurate and efficient detection of AFP-L3 is crucial for early HCC diagnosis and management.

Purpose of the Study:

• To develop and validate a chemiluminescent protein microarray for determining the AFP-L3/AFP index (AFP-L3%).
• To evaluate the diagnostic performance of the microarray assay compared to traditional ELISA methods.

Main Methods:

• A chemiluminescent protein microarray was designed using AFP-specific antibodies and Lens culinaris lectin on aldehyde-coated glass slides.
• Serum samples were analyzed for total AFP using ELISA and for AFP-L3 using a spin column pretreatment and ELISA.
• The AFP-L3% was calculated, and diagnostic performance metrics (sensitivity, specificity, ROC AUC) were assessed for both microarray and ELISA methods.

Main Results:

• The protein microarray demonstrated high sensitivity (89.74%) and specificity (100%) for HCC diagnosis at an AFP cut-off of 20 ng/ml.
• At an AFP-L3% cut-off of 0.1, the microarray showed 56.41% sensitivity and 100% specificity.
• The area under the ROC curve (AUC) for AFP was significantly higher (0.996) than for AFP-L3 (0.857) and AFP-L3% (0.827), indicating AFP's strong diagnostic power.

Conclusions:

• The developed chemiluminescent protein microarray is a sensitive, accurate, and efficient method for determining AFP-L3%.
• This assay holds promise for improving early screening and diagnosis of hepatocellular carcinoma.