The LRC- TriCEPS (Ligand-Receptor capture) applied to human primary T-cells

The LRC-TriCEPSTM technology to identify targets of peptides, proteins and antibodies on the living cells can also be applied to primary cells such as human T-cells. Further cell-cell interactions can be elucidated by identifying the binding targets of an extracellular domain of a membrane protein.

Protein level Volcano plot of LRC-TriCEPS

Figure 1: Protein level Volcano plot of LRC-TriCEPSTM workflow. CD28 was identified as the only target for anti-CD28 antibody and TRCB1 was identified as the target of anti-TRC2 α/β antibody. Proteins are considered significant candidates (red) if p-value<0.01 and log2FC>2.

To change as little as possible of the expressed proteins at the cell membrane (surfaceome) primary cells such as T-cells should be used freshly by the laboratory that isolated them. The LRC-TriCEPSTM technology has the advantage that the first part of the experiment can be conducted in any laboratory that is able to isolate primary T-cells. In a first step the protein/peptide ligand is coupled to TriCEPSTM (detailed manual provided by Dualsystems Biotech AG) and added to the freshly prepared primary T-cells. Then the cell pellets are frozen and shipped to Dualsystems Biotech AG. Dualsystems processes the cells, performs the LC-MS/MS measurements and data analyses and provides ca. 4-5 weeks later the results in form of a report. Here we show the volcano plot (Figure 1) that is used to display the data. It could be demonstrated that the two antibodies bind their expected targets in the cell membrane (CD28 and T-cell receptor beta chain 1(TRBC1)).

To show that the experiment can easily be performed by a collaboration of two laboratories we present here the results of an experiment that was conducted by Zellwerk GmbH in Germany and Dualsystems Biotech AG in Switzerland. Zellwerk GmbH isolated T-cells and coupled the ligands to TriCEPSTM. Zellwerk GmbH added then the TriCEPSTM coupled ligands to the cells, froze the cells and sent it to Dualsystems. In the presented experiment anti-CD28 antibodies were used as positive control and anti-TCR2alpha/beta antibodies were used as Ligand of Interest. The two expected targets CD28 and T-cell receptor beta chain 1(TRBC1) could be clearly identified.

For further information or to obtain a quote to perform a LRC-TriCEPSTM (Ligand-Receptor capture) experiments on primary cells please contact