Case study Dualsystems and Zellwerk GmbH
Example of LRC-TriCEPS applied to human primary T-cells

Figure 1: Protein level volcano plot of LRC-TriCEPS workflow. CD28 was identified as the only target for the anti-CD28 antibody and TRCB1 was identified as the target of the anti-TRC2 α/β antibody. Proteins are considered significant candidates (red) if p-value < 0.01 and log2 fold change >2.
To show that the experiment can easily be performed by a collaboration of two laboratories we present here the results of an experiment conducted by Zellwerk GmbH in Germany and Dualsystems Biotech AG in Switzerland. Zellwerk GmbH isolated T-cells and coupled the ligands to TriCEPS. Zellwerk GmbH added then the TriCEPS coupled ligands to the cells, froze the cells and sent the samples 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 LRC-TriCEPS (Ligand-Receptor capture) experiments on primary cells please contact triceps@dualsystems.com
The LRC-TriCEPS technology to identify targets of peptides, proteins and antibodies on 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.
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-TriCEPS technology has the advantage that the first part of the experiment can be conducted in any laboratory that can isolate primary T-cells. In a first step the protein/peptide ligand is coupled to TriCEPS – 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 the results approximately 4 – 5 weeks later in the form of a report. Here we show the volcano plot (Figure 1) used to display the data. It can be demonstrated that the two antibodies bind their expected targets in the cell membrane (CD28 and T-cell receptor beta chain 1 (TRBC1)).
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Customers Testimonials – LRC-TriCEPS Service
Testimonials from our customers who have used the LRC-TriCEPS technology – in collaboration with Dualsystems Biotech AG.
University Hospital of Lausanne (CHUV)
Thanks to their LRC-TriCEPS platform, we were able to identify a cellular factor playing a major role in Zika virus entry. This unique method was applied to living cells without the need for genetic modification. In our case, the method worked robustly across three different cell types. Notably, it enabled the identification of an unexpected target at the cell surface that would likely have been missed by standard screening approaches, as the protein lacks a classical membrane anchor. What impressed us most was that the implication of this target, discovered via their screening, was subsequently confirmed in our in vitro studies using different cellular models.
We greatly appreciated the collaboration with the DualSystems Biotech AG team. Their expertise, responsiveness, and willingness to support our project at every stage were key to its success. The data provided were of high quality and well presented, facilitating downstream interpretation and validation.
Milos Stojanov, PhD - Head of the Materno-fetal and Obstetrics Research Unit - University Hospital of Lausanne (CH)

Miloš Stojanov, PhD | Senior Lecturer (MER1)
Head of the Materno-fetal and Obstetrics Research Unit
Department Woman-mother-child (DFME)
1011 Lausanne
Tél: +41 21 314 05 17
OncoLille Cancer Institute

Best,
Silvia Gaggero, PhD
Mitra Lab, Inserm
OncoLille Cancer Institute
Lille, France
AstraZeneca

James Dodgson
AstraZeneca
Cambridge, UK.
UCF College of Medicine

Justine Tigno-Aranjuez, Ph.D.
Assistant Professor of Medicine
UCF College of Medicine
Cohbar

Dr. Lindsay Stark
Drug Discovery Scientist at CohBar
Technical University of Munich
Using LRC-TriCEPS, we aimed to identify novel direct cell surface receptors of our ligand of interest.
At any time, we experienced great support of Dualsystems Biotech. They kindly helped to find optimal conditions for our purposes and provided help with any kind of question before, during and after the experiment. LRC-TriCEPS allowed us to identify novel cell surface receptors of our ligand, which we could successfully validate in different cell types and with different biochemical assays. We can fully recommend Dualsystems Biotech and are looking forward to perform further analyses using LRC-TriCEPS.

Prof. Dr. rer. nat. Achim Krüger
Institute of Experimental Oncology and Therapy Research
Klinikum rechts der Isar, Technical University of Munich
University of Miyazaki

Hideyuki Sakoda, MD, PhD
Associate professor
Department of Biological Sciences, Faculty of Medicine, University of Miyazaki, Japan.
Lund University Diabetes Centre

Dr. Claire L. Lyons,
Associate Researcher
Unit of Medical Protein Science
Lund University Diabetes Centre
Sweden
Australian National University

The Australian National University
Co-Director, Centre for Personalised Immunology, NHMRC Centre of Research Excellence
College of Health & Medicine
The Australian National University
Harvard Medical School, Brigham and Women’s Hospital


Maximillian Rogers, PhD
Research Scientist
Harvard Medical School, Brigham and Women's Hospital
Department of Medicine, Cardiovascular Division
Boston, MA
Center for Biomolecular & Cellular Structure, Institute for Basic Science


Associate Professor
Graduate School of Medical Science and Engineering, KAIST
Chief Investigator
Center for Biomolecular & Cellular Structure, Institute for Basic Science (IBS)
Department of Internal Medicine Erasmus MC

Laboratory of Metabolism and Reproduction
Department of Internal Medicine
Erasmus MC
Rotterdam, The Netherlands
Seoul National University

Chung Hwan Cho, Ph. D. candidate
Environmental Health Microbiology Laboratory
Department of Environmental Public Health
Seoul National University
Immuno-Oncology Discovery from Bristol-Myers Squibb published in Nature
Identification of a new immune-oncology drug target using the LRC-TriCEPS platform on primary human T-cells.
The University of Oklahoma – Health Sciences Center

Anne Kasus-Jacobi, PhD
Associate Professor of Research
University of Oklahoma Health Sciences Center
Department of Pharmaceutical Sciences
Oklahoma City, Oklahoma, USA
CuroNZ Ltd

Frank Sieg, PhD
CSO
CuroNZ Ltd
Mangawhai in New Zealand
University of Pittsburgh

Maliha Zahid, M.D., Ph.D.
Assistant Professor
Departement of Developmental Biology
University of Pittsburgh
University of Oklahoma Health Sciences Center

Anne Kasus-Jacobi, PhD
Assistant Professor of Research
University of Oklahoma Health Sciences Center
Department of Pharmaceutical Sciences
Oklahoma City, Oklahoma, USA
Biomedical Research Institute
The identification of a T cell co-receptor for staphylococcal superantigens had been challenging due to the structural features of the interaction and its kinetics. However, working with Dualstystems Biotech AG, and with Dr. Paul Helbling in particular, and using the LRC-TriCEPS technology, we were able to identify a candidate that was subsequently corroborated by biochemical and functional assays. We are very happy with this collaboration , and sincerely recommend it for the identification of novel receptor or co-receptor candidates.

Chief Scientific Officer
Los Angeles Biomedical Research Institute at Harbor-UCLA Medical Center
Torrance, USA
QIMR Berghofer Medical Research Institute

Hepatic Fibrosis Group
QIMR Berghofer Medical Research Institute, Australia
University of Miami, Miller School of Medicine
I would like to thank once again the company and, particularly, Dr Helbling for his attention and collaboration.

Research associate professor
University of Miami, Miller School of Medicine
Department of Orthopedic surgery
Miami, Florida, USA
Münster University Hospital (UKM)

Working group from Prof. Dr. med. Luisa Klotz
Münster University Hospital (UKM), Germany
University of Manitoba

Associate Professor, Department of Human Anatomy and Cell Science
Associate Member, Spinal Cord Research Centre
Max Rady College of Medicine, Rady Faculty of Health Sciences
University of Manitoba
The Rockefeller University

Assistant Professor of Clinical Investigation
The Rockefeller University
Medizinische Hochschule Hannover
East Tennessee State University

Assistant Professor
East Tennessee State University
Igenica Biotherapeutics

Senior Director, Preclinical Development
Igenica Biotherapeutics
Centro de Estudos de Doenças Crónicas
« The fruitful collaboration with Dualsystems Biotech using the LRC-TriCEPS (CaptiRec) technology showed that even on insect cells receptors could be identified »

Principal Investigator at CEDOC
Centro de Estudos de Doenças Crónicas
Washington University School of Medicine
University of California San Francisco

Assistant Professor in Residence
University of California San Francisco (UCSF)
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Dualsystems Biotech AG
Dr. Paul Helbling
Grabenstrasse 11a
8952 Schlieren
Switzerland
phone +41 44 738 50 00
fax +41 44 738 50 05
LRC-TriCEPS Video
LRC-TriCEPS explained in a short video