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In Vitro Assays for Endolysin Efficacy and Safety Profiling
Nikolaos D Georgakis1, Panagiota D Pantiora1, Georgios E Premetis1
1Laboratory of Enzyme Technology, Department of Biotechnology, School of Applied Biology and Biotechnology, Agricultural University of Athens, 75 Lera Odos Street,, GR-11855, Athens, Greece.
Methods in Molecular Biology (Clifton, N.J.)
|August 7, 2026
Summary
This study presents three in vitro assays to test antimicrobial endolysins and their safety. These methods rapidly screen endolysin effectiveness and potential toxicity for new drug development.
Area of Science:
- Biochemistry
- Microbiology
- Drug Discovery
Background:
- Endolysins are promising antimicrobial agents.
- Evaluating their efficacy and safety is crucial for development.
- Standardized assays are needed for early-stage screening.
Purpose of the Study:
- To describe three complementary in vitro assays.
- To evaluate antimicrobial potential of endolysins.
- To assess cytotoxic safety of endolysins.
Main Methods:
- Kirby-Bauer disk diffusion for rapid screening of inhibition zones.
- Serial dilution assay for quantifying bactericidal efficacy (CFU reduction).
- Hemolytic assay using human red blood cells to assess cytotoxicity.
Main Results:
- The described assays provide a practical framework.
- Methods allow for rapid screening and dose-dependent assessment.
- Assays enable evaluation of both antimicrobial activity and host safety.
Conclusions:
- These complementary assays offer a reproducible method for early-stage endolysin screening.
- The assays facilitate efficient evaluation of antimicrobial potential and cytotoxic safety.
- This framework supports the development of novel endolysin-based therapeutics.
