A simple method to determine proteolytic activity of snake venoms

Area of Science:

• Biochemistry
• Toxicology
• Herpetology

Background:

• Snake venom characterization is crucial for antivenom development.
• Assessing proteolytic and hemorrhagic activities is vital for understanding venom toxicity.
• Existing methods can be complex and require significant animal use.

Purpose of the Study:

• To develop a simple, cost-effective method for assessing snake venom proteolytic activity.
• To correlate proteolytic activity with hemorrhagic activity.
• To evaluate the method's utility in assessing antivenom efficacy.

Main Methods:

• Radial hydrolysis assay on a gelatin-agarose plate to measure proteolytic activity.
• Determination of gelatinolytic dose (GD) for various snake venoms.
• Correlation analysis with hemorrhagic activity and assessment of EDTA inhibition.
• Application in evaluating antivenom neutralization capacity.

Main Results:

• A dose-response relationship was observed for all tested venoms.
• Proteolytic capacity (GD) ranged from 21 to 222 μg.
• A strong correlation (r² = 0.8774) was found between hydrolysis and hemorrhagic activity.
• EDTA partially inhibited hydrolytic activity.
• The method successfully evaluated antivenom neutralizing capacity.

Conclusions:

• The developed method is simple, cost-effective, and reliable for screening venom proteolytic activity.
• It can predict hemorrhagic activity and assess antivenom efficacy.
• This approach contributes to reducing animal usage in venom research.