Determination of trace alpha-fetoprotein variant by affinity adsorption solid substrate-room temperature...

Area of Science:

• Analytical Chemistry
• Biochemistry
• Materials Science

Background:

• Alpha-fetoprotein variant (AFP-V) is a biomarker requiring sensitive detection methods.
• Existing detection methods may lack the required sensitivity or specificity.
• Dual luminescent molecules offer unique properties for biosensing applications.

Purpose of the Study:

• To establish a new, highly sensitive method for trace AFP-V determination.
• To utilize the luminescent properties of a dendrimer-porphyrin conjugate for biosensing.
• To validate the method's accuracy using clinical serum samples.

Main Methods:

• Synthesis of a 3.5-generation dendrimer-porphyrin (3.5G-D-P) dual luminescent molecule.
• Labeling concanavalin A (Con A) with 3.5G-D-P to form 3.5G-D-P-Con A.
• Development of an affinity adsorption solid substrate-room temperature phosphorimetry (AA-SS-RTP) assay using polyamide membranes.
• Measuring the enhanced room temperature phosphorescence (RTP) signal correlated with AFP-V concentration.

Main Results:

• The 3.5G-D-P-Con A conjugate exhibited specific affinity adsorption for AFP-V.
• A sharp enhancement in RTP signal was observed upon AFP-V binding.
• The method demonstrated a linear correlation between the change in phosphorescence intensity (DeltaIp) and AFP-V content.
• High sensitivity and accuracy were achieved for trace AFP-V detection in serum samples.
• Results showed good agreement with the established ELISA method.

Conclusions:

• The developed AA-SS-RTP method provides a sensitive and accurate approach for quantifying trace AFP-V.
• The 3.5G-D-P-Con A conjugate is a promising tool for AFP-V detection.
• This technique holds potential for clinical diagnostics and biomarker monitoring.