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LRC-TriCEPS Service

LRC-TriCEPS™


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-next generation
  • 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

Decoding the extracellular interactome using LRC-TriCEPS™

A novel approach to analyse 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 the ETH Zurich who published the technology in Nature Biotechnology (Frei et al.(2012) Nature Biotechnology 30:997-1001).

Orphan ligands:

  • Extracellular proteins
  • Peptide ligands
  • Antibodies
  • Viruses
  • Engineered affinity binders

Dualsystems Biotech AG, is a biotech company that provides services to analyse protein-protein interactions.

You are looking for the targets of your ligand (peptide, protein, antibody, and virus) that are relevant in the living organism? Give us a call +41 44 738 50 00 or fill in the form on the right side and we get in contact to see how the LRC-TriCEPS technology (ligand-receptor capture) can answer your question.


Ligand-receptor capture

LRC-TriCEPS™ (CaptiRec) is a novel approach which enables the identification of cell surface receptors and off-targets on the living cells for a wide range of orphan ligands, such as:

  • Peptides
  • Protein
  • Antibodies
  • Engineered affinity binders
  • Viruses

Workflow-2016-April

Ligand-receptor-Flyer-Dualsystems Biotech-2017-PDF

Availability

The TriCEPS™-based ligand-receptor capture technology is now available.

Principle

Principle of LRC-TriCEPS™-based 

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

LRC-TriCEPS™ – a trifunctional chemoproteomics reagentReceptor-mit-Ligand-Ausschnitt-2016-small

TriCEPS™ structure:

  • NHS-ester for attachment to the ligand
  • Protected hydrazine function for capturing the interacting receptor
  • Biotin function for purification of ligand-receptor peptides

Requirements

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)
Trastuzumab (Herceptin) Breast cancer tissue ErbB2
Ankyrin repeat proteins BT-474 human breast
cancer cells
ErbB2, domain I
Vaccinia virus HeLa CCL2 cells AXL, M6PR, DAG1, CSPG4

From Frei et al. (2012) Nature Biotechnology 30:997

Publications

Company

About Dualsystems Biotech


Dualsystems Biotech Schlieren Zurich Switzerland

Dualsystems Biotech is a provider of proteomics services for industry and academia. The LRC TriCEPS technology platform is designed to identify targets and off-targets irected to identify intracellular as well as extracellular drug targets. Our specialities LRC-TriCEPS™, ligand-receptor capture and FLOW-TriCEPS™ make use of mass spectrometry analysis of protein complexes.

  • Established in 2000
  • Facilities in Zurich, Switzerland
  • Provider of target screening technologies
  • Swiss Quality

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