Now available: The next generation of LCR-TriCEPS
TriCEPSTM V.3.0 enables receptor identification for your orphan ligands at the surface of living cells without genetic manipulation. Key features of the new TriCEPSTM V.3.0 and LRC- TriCEPSTM technology are:
- Reduction of the number of cells needed:
- TRICEPS V3.0 LRC experiments require 5-10 fold less starting material for successful receptor identification compared to TRICEPS V2.0.
- High coverage of the surfaceome.
- A modified LRC workflow enabled by TriCEPSTM V.3.0 allows for the theoretical identification of up to 85% of all putative cell surface proteins.
- Identification of N-, C-, O- glycosylated targets
- More sensitive identification of low copy number cell surface receptors through all tryptic peptides.
Decoding the extracellular interactome using LRC-TriCEPS™
A novel approach to the analysis of extracellular cell signaling by discovering cell surface receptors and off-targets for a ligand of interest on the living cell: label ligand with the LRC-TriCEPS™crosslinker, incubate with living cells or tissue expressing target receptor(s), purify receptor crosslinked peptides, analyse by LC-MS/MS. This new technology was invented by Andreas Frei and Bernd Wollscheid from ETH Zurich who published the technology in Nature Biotechnology (Frei et al.(2012) Nature Biotechnology 30:997-1001).
- Extracellular proteins
- Peptide ligands
- Engineered affinity binders
Dualsystems Biotech AG is a biotech company providing analytic services for protein-protein interactions.
If you are looking for the targets of your ligand – peptide, protein, antibody, and virus – that are relevant in the living organism, call us on +41 44 738 50 00 or fill in the form on the right hand We will contact you to see how LRC-TriCEPS technology (ligand-receptor capture) can answer your questions.
LRC-TriCEPS™ (CaptiRec) is a novel approach enabling the identification of cell surface receptors and off-targets on living cells for a wide range of orphan ligands, such as:
- Engineered affinity binders
The TriCEPS™-based ligand-receptor capture technology is now available.
LRC-TriCEPS™ (CaptiRec) is a fast and sensitive approach to discover cell surface N-glycosylated receptors for a ligand of interest:
- Label ligand with the TriCEPS™ chemoproteomic reagent
- Activate target cells or native tissue to oxidize cell surface glycans
- Incubate ligand-TriCEPS™ complex with activated cells
- TriCEPS™ immediately links ligand and cognate receptor(s)
- Isolate all membrane proteins
- Tryptic digest of all membrane proteins
- TriCEPS™-mediated purification of N-glycosylated peptides
- Deglycosylation of peptides
- Identification of peptides by mass spectrometry
- Analysis of data using statistics and bioinformatics
- NHS-ester for attachment to the ligand
- Protected hydrazine function for capturing the interacting receptor
- Biotin function for purification of ligand-receptor peptides
Requirements for one LRC experiment: Ligand in triplicate compared to control in triplicate
- 300 µg ligand containing a free amino group
- 600 Mio cells or 50 µm tissue slice
Examples of successful ligand-receptor capture (LRC) identifications
|Ligand||Target cells or tissue||Receptors identified|
|Insulin||Murine adipocytes, Jurkat cells||Insulin receptor|
|Apelin-17||U2OS osteosarcoma cells||Apelin receptor (GPCR)|
|Ankyrin repeat proteins||BT-474 human breast
|ErbB2, domain I|
|Vaccinia virus||HeLa CCL2 cells||AXL, M6PR, DAG1, CSPG4|
- Li Y, Ozment T, Wright GL, Peterson JM. PLoS One. 2016 Oct 11;11 Identification of Putative Receptors for the Novel Adipokine CTRP3 Using Ligand-Receptor Capture Technology
- Manish P. Ponda and Jan L. Breslowa, Serum stimulation of CCR7 chemotaxis due to coagulation factor XIIa-dependent production of high-molecular-weight kininogen domain 5
- Heather L. Glasgow, Michael A. Whitney, Larry A. Gross, Beth Friedman, Stephen R. Adams, Jessica L. Crisp, Timon Hussain, Andreas P. Frei, Karel Novy, Bernd Wollscheid, Quyen T. Nguyen, and Roger Y. Tsien – Laminin targeting of a peripheral nerve-highlighting peptide enables degenerated nerve visualization, (2016) Glasgow et al. 10.1073/pnas.1611642113
- Jonathan M Peterson, April 2016, The FASEB Journal, vol. 30 no. 1 Supplement 1249.2 Identification of cell surface receptors for the novel adipokine CTRP3
- Alisson M. Gontijo, 29 October 2015 Nature Communications 6, Dilp8 requires the neuronal relaxin receptor Lgr3 to couple growth to developmental timing
- Damaris Bausch-Fluck et al. (April 2015) A Mass Spectrometric-Derived Cell Surface Protein Atlas
- Frei et al. (2012) Nature Biotechnology 30:997-1001
- Frei et al. (2013) Nature Protocols 8:1321-1336
- Slavoff and Sagathelian (2012) Nature Biotechnology 30:959-961
- De Souza (2012) Nature Methods 9, 1044–1045
- Donner (2012) Nature Chemical Biology 8:950
- Protter data base: Omasits U, Ahrens CH, Müller S, Wollscheid B. Bioinformatics. 2013 Nov21.