Activity Detail
Seminar
New advances in proteomic analyses
Felix Elortza
At Proteomics Platform have been settling different methodologies to improve
our proteomic analysis performance in terms of sensitivity, throughput, and
quality standards.We are all aware that in biomedicine and many basic research
projects starting with less material is more. Biopsies can be smaller, required cell
amount can be less and therefore experiments can be cheaper, etc. By applying
the so called SP3 digestion method we are now able to digest very scarce starting
material samples being our proven limit the human oocyte analysis. In
proteomics one of the most demanded outcomes is to identify as much as
possible proteins, detect which ones are differentially expressed and to quantify
these differences. We have been working to implement the Dia NN (Data
Independent Acquisition Neural Network) method within our pipeline. Overall,
this method helps identifying more peptides and quantifying more proteins in
complex mixtures. Finally, increasing throughput is also a needed to face some
biomedical studies. We have settled a microtiter like PVDF based approach to
reduce, alkylate and perform the tryptic digestion over 96x4 samples in 2 3 days
of work. These improvements, together with the EVOSEP One chromatographer
couple on line to a TIMS Tof Pro mass spectrometer, make our pipelines capable
to tackle high sensitivity and high throughput projects. Different examples of
projects where these methods have been applied will be shown.