Development of a Methodology Based on Optical Interferometry for Measuring Fibrinolytic Activity

Area of Science:

• Biomedical Engineering
• Biochemistry
• Analytical Chemistry

Background:

• Fibrinolytic activity is crucial for cardiovascular disease management and fibrinolytic drug development.
• Current assays for fibrinolysis lack real-time, label-free capabilities.
• Accurate measurement of fibrinolysis is essential for diagnosis and treatment.

Purpose of the Study:

• To develop a novel, real-time, label-free method for dynamic detection of fibrinolytic activity.
• To utilize ordered porous layer interferometry (OPLI) for sensitive fibrinolysis measurement.
• To establish a more targeted in vitro research method for fibrinolysis.

Main Methods:

• Constructed a fibrinolytic response interference layer using silica colloidal crystal (SCC) films loaded with fibrin and plasminogen (Plg).
• Employed OPLI to track real-time optical thickness changes (ΔOT) caused by fibrinolysis.
• Optimized Plg content and experimental parameters for the OPLI system.

Main Results:

• Demonstrated high sensitivity and wide linear ranges for lumbrokinase (12-6000 U/mL) and streptokinase (10-2000 U/mL).
• Achieved lower detection limits and higher linearity compared to the traditional fibrin plate method.
• Successfully recorded the kinetic process of fibrinolysis and calculated kinetic parameters.
• Showed minimal interference from other blood proteins and reliability in whole blood samples.

Conclusions:

• The OPLI-based assay provides a sensitive, real-time, and label-free method for dynamic fibrinolytic activity detection.
• This technique offers advantages over traditional methods, including improved linearity and lower detection limits.
• The developed assay is reliable for analyzing fibrinolytic activity in whole blood, aiding cardiovascular disease research and drug development.