Protein-protein interactions
Proteins are the fundamental building blocks and control all biological systems in a cell. The physical interaction between proteins define specific biological functions and signaling networks.
Explore how our customers have used Evosep One to scale the study of complex protein environments and understand cell biology.
SCALABLE PROTEIN-PROTEIN INTERACTION ANALYSIS TO UNDERSTAND DISEASE
Proteins are the functional molecules in our body and contribute to all processes of life. Like humans, proteins act in social networks and the understanding of the cellular machinery and disease mechanisms can be found in the dynamic networks of protein complexes. Insights into the physical rewiring of protein-protein interactions in response to i.e., mutations and pathological conditions can lead to the identification of potential druggable targets.
By using quantitative mass spectrometry-based proteomics, unbiased disease-mediated changes in protein-protein interactions can be studied on a global scale. The Evosep One nicely facilitates large-scale studies in a high-throughput manner.
PARTIAL ELUTION ON EVOTIPS ARE CRUCIAL FOR INTERACTOME STUDIES
It is important to leave the proteins in their native conformation for pull-down assays. Thus, non-ionic detergents are often used in the lysis buffer to solubilize proteins.
However, even minute levels of detergent severely decreases chromatographic resolution in LC-MS as it accumulates in the analytical column and the backpressure increases with each sample.
This can be circumvented by using the Evosep One as the Evotip is a disposable trap column, which is discarded after each sample. Our patented partial elution concept leaves all contaminants on the tip ensuring the feasibility of large-scale interactome studies.
LARGE-SCALE MAPPING OF PROTEIN NETWORKS IN THE LUNGS
Researchers from the Olsen Group at University of Copenhagen have mapped the EGF-dependent in vivo phosphotyrosine signaling in lung tissue quantifying more than 1000 phosphotyrosine sites.
To assign function to all EGF-regulated sites, they determined their recruited protein signaling complexes in lung tissue by interaction proteomics. This provides insights to how mutations near tyrosine residues introduce molecular switches that rewire cancer signaling networks and revealed oncogenic properties of such a lung cancer EGFR mutant.
They demonstrated the scalability of their approach by performing more than 1000 phosphopeptide pulldowns and analyzed them with the 60 samples per day method using DIA on a Q Exactive HF-X mass spectrometer.
Read publication here
OPEN CELL PROVIDES A NEW VIEW ON HUMAN CELLULAR ORGANIZATION
The collaboration between the Mann group at the Max-Planck Institute for Biochemistry and the Chan Zuckerberg Biohub integrates genome engineering, confocal imaging, mass spectrometry and data science to systematically map protein localization in live cells and protein interactions under endogenous expression conditions.
They generated 1311 CRISPR-edited cell lines and applied a machine learning framework to encode the interaction and localization profiles of each protein. This resulted in almost 4000 pull-downs, which were analyzed with the 60 samples per day method, in just over two months of measurement time on a timsTOF Pro.
Their approach provides a data-driven description of the molecular and spatial networks that organize the human proteome.
Read publication here
EVOSEP WEBINAR
MEET OUR USERS and their research with protein-protein interaction analysis
ROBUST AND COMPREHENSIVE PROTEIN-PROTEIN INTERACTION ANALYSIS WITH EVOSEP ONE
Available on demand
During this webinar Leonard Foster (University of British Colombia, Vancouver) and Lisa Jones (University of Maryland, Baltimore) will provide their insight into the use of Evosep One in the study of complex protein environments.
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Evosep One solves the proteomics dilemma – covering high throughput analysis and proteome depth
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end-to-end, fully automated digestion protocol and evotip pure workflow on the opentrons ot-2
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fully automated, rapid, robust sample loading on evotip pure with the agilent assaymap bravo
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EVOSEP ONE: A Standardized Separation
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