Screening Platform Based on Robolid Microplate for Immobilized Enzyme-Based Assays.

Area of Science:

• Biochemistry
• Analytical Chemistry
• Biotechnology

Background:

• Enzyme-based assays are crucial for biochemical analysis and disease diagnosis.
• High-throughput screening methods are needed to accelerate the analysis of multiple analytes.
• Current methods may lack efficiency, cost-effectiveness, or throughput for broad application.

Purpose of the Study:

• To develop a facile, cost-effective, and high-throughput screening method for enzyme-based assays.
• To establish a novel Robolid/Microplate (RLMP) platform for rapid biochemical analysis.
• To demonstrate the platform's utility in detecting clinical markers.

Main Methods:

• Enzyme immobilization onto commercial robolids.
• Integration of enzyme-immobilized robolids with standard 96-well microplates.
• Initiation and quenching of enzymatic reactions via sandwiching/unsealing of robolids and microplates.
• Utilizing urea as a model analyte for validation.

Main Results:

• The RLMP platform demonstrated a large linear detection range (up to 10 mM for urea).
• Achieved a fast analysis time of 30 minutes for 96 samples.
• Exhibited good reproducibility, stability, and a strong correlation with a commercial urea test kit.
• Successfully measured urea in human urine and serum samples.

Conclusions:

• The developed RLMP platform offers a promising solution for high-throughput enzyme-based assays.
• The platform facilitates simultaneous measurement of multiple analytes with independent analysis time.
• It shows significant potential for clinical marker detection, disease diagnosis, and broader biochemical analysis.