Evaluating the toxicity of proteins without using animals: an ambitious goal, but achieved by the Protox project. This pioneering initiative, coordinated by the French laboratory ANSES Fougèresmarks a decisive step towards a more ethical and sustainable future, exploiting advanced methodologies such as in silico and in vitro tests.
Thinking about toxicity tests without using animals seemed, until a few years ago, a distant goal. With the project ProtoxHowever, change has become reality. Launched in December 2022 and concluded in May 2024, Protox was developed by the ANSES Fougères laboratory, in collaboration with the Luxembourg Institute of Science and Technology, in response to a request from EFSA, the European Food Safety Authority.
Kevin Hogeveentoxicologist and project coordinator, explains:
The objective was simple in its complexity: to guarantee a safe evaluation of proteins, without the use of animals, using innovative and ethically respectful methodologies.
In addition to an ethical motivation, the project sought to respond to a regulatory need: traditional animal-based methodologies are increasingly under scrutiny for their scientific and moral limitations.
A method based on computers and cell cultures
Protox’s strong point is the adoption of a gradual strategydesigned to completely replace animal models. First, they were analyzed 7,000 potentially toxic proteinsdivided according to their toxicity mechanisms. From here, a two-step methodology was developed:
- In silico analysis: In this first phase, the computer plays the main role. The structures of the proteins are analyzed and compared with the toxic ones already known, identifying similarities that may indicate a risk. If the protein comes from species known to produce toxic substances, further attention is focused on it.
- In vitro tests: The second phase takes place in the laboratory, using cell cultures to simulate the effects of proteins on different tissues. If exposure occurs orally, for example, the effects on the intestine are evaluated. If the protein shows the ability to cross the intestinal barrier, tests are conducted on internal organs such as the heart or brain. All this happens without ever resorting to animal testing.
Because Protox is a revolution for food safety
Protox’s importance is not limited to eliminating animal testing. This project represents a new scientific standardwith a positive impact on both research ethics and food safety.
With climate change modifying natural balances, more and more toxic proteins, produced by fungi, algae or plants, risk contaminating our foods. Protox offers a quick and effective system to identify these risks without sacrificing animals. Hogeveen adds:
We wanted to propose a model that was not only ethical, but also accessible. In vitro and in silico techniques can be easily adopted by laboratories around the world.
Protox’s final report, approved by EFSA, was published on 5 November 2024. Although the method is not yet integrated into European regulations, it will be used alongside traditional tests to evaluate its comparative effectiveness. This project, however, has already demonstrated that a science without animals is possiblemarking an important step towards more sustainable and compassionate research.
