CovalX Complex Analysis
This website uses cookies so that we can provide you with the best user experience possible. Cookie information is stored in your browser and performs functions such as recognising you when you return to our website and helping our team to understand which sections of the website you find most interesting and useful.
Protein Interaction Analysis
Talk to Our Scientists
CovalX has developed and optimized a method for a fast, sensitive and accurate analysis of protein complexes directly by High-Mass MALDI mass spectrometry. Unfragmented and undigested, the protein complexes are detected intact using specially developed cross-linking reagents and high-mass detection system. There is no sample pre-labeling or tagging required and no immobilization, the samples are measured directly in solution.
Using cross-linking chemistry and High-Mass MALDI ToF mass spectrometry, it becomes possible to easily analyze non-covalent protein complexes in the 1-2000 kDa range. The sample to be analyzed can be a simple aggregate multimers (dimer, trimer, etc), antibody fragment (Fab: one-to-one), monoclonal (two-to-one), or more advanced stoichiometry of nearly any protein-protein interaction.
The use of High-Mass MALDI allows a much faster, cheaper and more accurate method for studying protein interactions:
- The stoichiometry of the complex: unique information for understanding the shape of your protein complex.
- Clear identification of the partners involved in the complex.
- No need for immobilization, the complex is detected without labeling.
- Fast: 30 minutes per assay.
- Specific: only the specific complexes are detected.
- Sensitive: Only a few hundred femtomoles of the protein sample is a few µl are required to achieve the analysis.
In the spectrum presented, the protein complex AMPk is analyzed by High-Mass MALDI ToF mass spectrometry before cross-linking (black) and after cross-linking (red) with CovalX’s K200 MALDI MS Analysis kit. (Contact Us to learn more about this Kit) Before cross-linking, the three subunits α, β, and γ of the protein complex are detected with m=30.22 kDa, m=37.38 kDa, and m=64.97 kDa. After cross-linking, the intact protein complex corresponding to the stoichiometry [α·β·γ] is detected with m=129.95 kDa. Riek U et al.; J. Biol. Chem.; 2008.
CovalX services then use our Complex Tracker software for the internal correction and direct overlay and subtraction analysis of the control and complex spectra providing a rapid and accurate characterization of the complex.