A recent study shows a promising future for non-animal testing. In June of 2015, a group of researchers from the University of Torino, residing in Italy, went public with their findings. “Electrochemistry of Canis familiaris cytochrome P450 2D15 with gold nanoparticles: An alternative to animal testing in drug discovery” has opened the door for more modern and more accurate findings to follow.
This research involves the extensive study of the Canis familiaris cytochrome P450 2D15, a specific portion of the canine’s cytochromes, located in the inner membrane of the mitochondria. According to the study, Canis familiaris is “one of the most widely studied animal models used in safety determination of new pharmaceuticals” (110). Typically the canine hepatic cytochromes P450 are targeted to test the toxicological effects of pharmaceuticals, even though they are only about 75% similar to human cytochromes. Although this number seems reasonable for comparison, wouldn’t it be comforting to compare cytochromes with 100% identicality?
These researchers report the first ever recombinant expression and purification of Canis familiaris by first amplifying liver cDNA from a canine subject, cloning it into an expression vector, and then fusing with a protein to form ‘native P450 enzymes’. Once purified through isolation methods, the P450 2D15 enzyme was immobilized on a glassy carbon. They performed this through two techniques:
- Entrapping the enzyme using specific volumes of proteins and surfactant solutions before dropping it on the glassy carbon.
- Immobilizing the enzyme using DDAB (didoeclyimethylammonium bromide) stabilized gold nanoparticles.
The purpose of these materials is to perform electrocatalysis experiments using chronoamperometry. The results identified all mutations of the Canis familiaris liver cDNA. According to their results, “These values are in the range of other electrochemically determined midpoint potentials reported in the literature for different human P450 enzymes” ( 113).
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This new study contains a lot of vocabulary that is difficult to absorb, however the development of this electrochemical platform can be used to screen animal cytochromes, which is a step in the right direction for reducing the need for animal testing. Through this new platform there are higher levels of predictability and accuracy in the animal models. Once a cell is isolated and purified, they can test a desired drug directly on the cell, and further analysis can determine a detailed list of mutations within that exposed cell.
According to their conclusions, this method of pharmaceutical testing “brings us a step closer to the realization of an in vitro electrochemical toxicity screening, an alternative to in vivo animal testing and sacrifice” (115).
Another possible outcome of this new technology includes the additional cloning of these purified cells, which can lead to further reduction of animal subjects. Also, examination of the diversity between human and animal cells can give rise to a more reliable interpretation of the data introduced (not just when comparing canines to humans).
If successful, we can undoubtedly say whether or not the effects of an animal will be the same as in human trials. Thanks to the advancements in technology, we will no longer question the level of diversity between species. Hopefully, the differences between beings will be seen as too great for researchers to even consider animal testing.