Initially, this technique was developed for fundamental research. Methods developed in this context being now sufficiently robust and proven, Phylogene undertook to adapt it to the analysis of large molecules. To this end, the MS Phylogene platform uses an original configuration of mass spectrometry coupled to high pressure liquid nano-chromatography (nanoLC-MS/MS).
Quantitative measurement of proteins
The objective is to achieve a quantitative comparison of all the proteins in two complex samples. In concrete terms, samples of skin explants, reconstituted skin, D-Squame, fibroblasts, keratinocytes or other biopsies after being subjected to an active are compared with their placebo control. The platform is powerful enough to be able to identify 200 to 2000 different proteins per sample and measure their relative amounts. Comparing this data with the data coming from the placebo gives a first indication on the possible (and sometimes unexpected) effects of molecules studied.
“The platform is of great interest for applications in cosmetics,” explained Gilbert Skorski, President of Phylogene. “Proteins are, indeed, the primary players in the cell’s function and structure.” Hence the value of knowing the impact of a formula or of an active on these proteins, especially with the current ban on animal testing. “Techniques looking for activated genes move into the background, because molecules expressed from the genes are not directly identified by these techniques,” he added.
Another advantage put forward by Phylogene is the quantitative nature of the technique. “We get immediate information on the impact and intensity of the resulting phenomena,” emphasised Skorski. The information of course, needs to be analysed and interpreted afterwards.
Finally, the fact that this technique is not targeted enables to not limit research to preconceived ideas that one may have on the effect of an active, but on the contrary to understand what is happening globally, positively or negatively. “This avoids having to deal with the extra costs relative to the late discovery of unexpected negative effects of an active, which was interesting beforehand.” Conversely, good surprises can also emerge just like unexpected effects, yet of particular interest.
All this, according to Phylogene, in less than a month’s time for a complete screening of the active.
Analysis of effects
Once that step is completed, it is possible to perform a bio-informatic and statistical analysis to identify the metabolic pathways and the activated biological or molecular functions. Phylogene has developed for this purpose the Coravalid™ module.
“This step is extremely important, because given the mass of information, without these additional analyses, experience shows that people would tend to only focus on what they already know and miss the new things corresponding to unexpected effects. There is a serious risk of losing the benefit of the relative quantification of these proteins,” explained Skorski. “Finally it is also possible to target multiple proteins, identified as biomarkers and thus to monitor the specific effect of the active, or to perform the same approaches on metabolites generated by these proteins, or to study the post-translational changes of the proteins,” he concluded.