Fibrin plate method with reagents purified by affinity chromatography and its use for determination of fibrinolytic and other proteolytic activity in saliva, bile and plasma

Area of Science:

• Biochemistry
• Analytical Chemistry
• Biotechnology

Background:

• The fibrin plate method is a standard technique for assessing fibrinolytic activity.
• Accurate quantification of plasmin and plasminogen activators is crucial in various biological and clinical contexts.
• Existing methods may have limitations in sensitivity or require larger sample volumes.

Purpose of the Study:

• To present a modified fibrin plate method for estimating plasmin and plasminogen activator activity.
• To improve the sensitivity and reproducibility of fibrinolytic activity assays.
• To demonstrate the applicability of the modified method in analyzing biological fluids like bile and saliva.

Main Methods:

• A modified fibrin plate method was developed using plasminogen-free human fibrinogen and purified plasminogen.
• Agarose was employed as a stabilizing medium in the fibrin plates.
• Fibrin plates were prepared with and without a constant amount of plasminogen to differentiate between plasmin and plasminogen activator activities.
• The method was validated using sterile bile and saliva samples.

Main Results:

• The modified fibrin plate method demonstrated sensitivity in detecting plasmin and plasminogen activator activity.
• Activator activity was successfully identified in sterile bile and saliva samples.
• The assay requires small volumes of reagents and samples, indicating efficiency.
• The method exhibited low error rates and good reproducibility.

Conclusions:

• The modified fibrin plate method offers a sensitive, reproducible, and efficient approach for quantifying plasmin and plasminogen activator activity.
• This technique is suitable for analyzing biological samples, including bile and saliva.
• The method provides a valuable tool for research in fibrinolysis and related fields.