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.
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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)).
Customers Testimonials – LRC-TriCEPS Service
Testimonials from our customers who have used the LRC-TriCEPS technology – in collaboration with Dualsystems Biotech AG.
UCF College of Medicine
“DualSystems LRC-TriCEPS platform helped us in the identification of a novel allergen receptor. This finding was central to our recently published work and funded grant! DualSystems was very helpful during the entire process, if you are considering using this platform, do not hesitate. The process is straightforward and the support is excellent!”
Justine Tigno-Aranjuez, Ph.D.
Assistant Professor of Medicine
UCF College of Medicine
Justine Tigno-Aranjuez, Ph.D.
Assistant Professor of Medicine
UCF College of Medicine
2023-03-16T06:40:12+00:00
https://www.dualsystems.com/testimonials/ucf-college-of-medicine/
Cohbar
DualSystems' LRC-TriCEPS platform was exactly what we needed to identify potential targets and guide us in mechanism of action studies for our peptide program. They helped us every step of the way -- from understanding their capabilities, to guiding study design, and supporting sample preparation. The report they delivered exceeded our expectations and truly stands out from data reports we've received from other companies. They were readily available to answer questions and discuss concerns or outcomes. We are very encouraged by the results and are currently following up on the targets identified. We highly recommend working with DualSystems and incorporating the LRC-TriCEPS platform into your program. We look forward to working with the DualSystems team again in the future.
Dr. Lindsay Stark
Drug Discovery Scientist at CohBar
Dr. Lindsay Stark
Drug Discovery Scientist at CohBar
2023-01-18T08:14:48+00:00
https://www.dualsystems.com/testimonials/cohbar/
Technical University of Munich
Technical University of Munich – Institute of Experimental Oncology and Therapy Research
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
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
2022-07-27T14:23:02+00:00
https://www.dualsystems.com/testimonials/technical-university-of-munich/
University of Miyazaki
We appreciate the support of Dualsystems Biotech and their LRC-TriCEPS technology we were able to identify a binding protein for our insulinotropic peptide
Hideyuki Sakoda, MD, PhD
Associate professor
Department of Biological Sciences, Faculty of Medicine, University of Miyazaki, Japan.
Hideyuki Sakoda, MD, PhD
Associate professor
Department of Biological Sciences, Faculty of Medicine, University of Miyazaki, Japan.
2022-04-19T11:45:22+00:00
https://www.dualsystems.com/testimonials/university-of-miyazaki/
Lund University Diabetes Centre
We wanted to use LRC-TriCEPS to characterise the mechanism of action of our ligand of interest. Not only were several candidate targets identified but we have successfully validated two of these receptors in insulin producing beta cells. DualSystems Biotech were very helpful and involved from study design, to experiments and reporting. Even after the initial report, DualSystems Biotech have answered follow up queries and provided assistance regarding publications. I highly recommend the LRC-TriCEPS technology and DualSystems Biotech and Paul Helbling and his team to work with.
Dr. Claire L. Lyons,
Associate Researcher
Unit of Medical Protein Science
Lund University Diabetes Centre
Sweden
Dr. Claire L. Lyons,
Associate Researcher
Unit of Medical Protein Science
Lund University Diabetes Centre
Sweden
2021-10-25T05:58:59+00:00
https://www.dualsystems.com/testimonials/lund-university-diabetes-centre/
Australian National University
“We had a challenging project where we aimed to discover the receptor of our ligand of interest in primary human cells. Dualsystems Biotech took the challenge and provided outstanding support throughout the entire process, from the experimental design to data analysis. Using their LRC-TriCEPS technology, we could identify a candidate receptor and additional downstream interacting partners that we have now validated. We look forward to new collaborations and discoveries with Dr Paul Helbling and his team”. – Prof Carola Vinuesa, ANU
The Australian National University
Co-Director, Centre for Personalised Immunology, NHMRC Centre of Research Excellence
College of Health & Medicine
The 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
2021-01-11T08:16:02+00:00
https://www.dualsystems.com/testimonials/australian-national-university/
Harvard Medical School, Brigham and Women’s Hospital
We used DualSystems to help identify a potential novel receptor for a ligand that despite being studied for decades had unclear catabolism mechanisms involving its receptor mediated uptake. The DualSystems team will walk you through the process, including early validation of your ligand being used for this technology, data collection and analysis, and provide an easy to follow report on the findings. For anyone interested in receptor-ligand interactions, DualSystems and the TriCEPS approach is worth strongly considering involving in your research program, as it can quickly help identify receptors that would have been very difficult to do so without.
Maximillian Rogers, PhD
Research Scientist
Harvard Medical School, Brigham and Women's Hospital
Department of Medicine, Cardiovascular Division
Boston, MA
Maximillian Rogers, PhD
Research Scientist
Harvard Medical School, Brigham and Women's Hospital
Department of Medicine, Cardiovascular Division
Boston, MA
2020-11-09T13:27:52+00:00
https://www.dualsystems.com/testimonials/harvard-medical-school-brigham-and-womens-hospital/
Center for Biomolecular & Cellular Structure, Institute for Basic Science
“It was my great pleasure to collaborate with Dualsystems Biotech. We have been tested various proteomic analysis to identify the specific receptor for our secreted ligand. But, only LRC-TriCEPS experiment successfully gave us a right answer, because this receptor and ligand interactions were indeed relatively weaker than other general receptor and ligand interactions. The team very kindly consulted the whole experimental steps from the experimental design to the final data analysis. We are now going to use TriCEPS again for several other secreted ligands with a great hope for discovery of new receptors. Best Regards, Ho Min Kim”
Associate Professor
Graduate School of Medical Science and Engineering, KAIST
Chief Investigator
Center for Biomolecular & Cellular Structure, Institute for Basic Science (IBS)
Associate Professor
Graduate School of Medical Science and Engineering, KAIST
Chief Investigator
Center for Biomolecular & Cellular Structure, Institute for Basic Science (IBS)
2020-08-12T13:02:47+00:00
https://www.dualsystems.com/testimonials/center-for-biomolecular-cellular-structure-institute-for-basic-science/
Department of Internal Medicine Erasmus MC
We were fortunate to partner with Dualsystems Biotech AG to discover cell-surface targets for the short-peptide ligand we had been working on for several years. We decided to try the LRC-TriCEPS approach to identify cell-surface binding partners for the ligand after several earlier aborted attempts using different techniques. Using LRC-TriCEPS we have identified targets that we have now been able to verify functionally and biochemically using other methods. It was a pleasure to work with Dr Paul Helbling and his team who were extremely helpful guiding us through the experimental process and assisting with analysis of the data. We were impressed by the quality of the data and the ease with which we were able to run the study.
Dr Patric Delhanty
Laboratory of Metabolism and Reproduction
Department of Internal Medicine
Erasmus MC
Rotterdam, The Netherlands
Dr Patric DelhantyLaboratory of Metabolism and Reproduction
Department of Internal Medicine
Erasmus MC
Rotterdam, The Netherlands
2020-04-28T11:54:52+00:00
https://www.dualsystems.com/testimonials/department-of-internal-medicine-erasmus-mc/
Seoul National University
"I have used many binding assays and was in the verge of giving up. Then I have contacted Dr. Paul Helbling and his group. The LRC-TriCEPS experiment was effective and let us go over the obstacle we had during the project. We look forward to using TriCEPS in the future".
Chung Hwan Cho, Ph. D. candidate
Environmental Health Microbiology Laboratory
Department of Environmental Public Health
Seoul National University
Chung Hwan Cho, Ph. D. candidate
Environmental Health Microbiology Laboratory
Department of Environmental Public Health
Seoul National University
2020-01-29T14:05:52+00:00
https://www.dualsystems.com/testimonials/seoul-national-university/
The University of Oklahoma – Health Sciences Center
I was very pleased with my collaboration with the team at Dualsystems. We had to work with difficult cells and the team helped us develop modifications to the original protocol to be able to use the TriCEPS approach with our cells. The whole team was very responsive, involved, and knowledgeable and this led to a successful identification of a receptor for our ligand of interest. We have since then confirmed the interaction and we are now getting ready to publish this study, which will have implications in the treatment of ocular injuries.
Anne Kasus-Jacobi, PhD
Associate Professor of Research
University of Oklahoma Health Sciences Center
Department of Pharmaceutical Sciences
Oklahoma City, Oklahoma, USA
Anne Kasus-Jacobi, PhD
Associate Professor of Research
University of Oklahoma Health Sciences Center
Department of Pharmaceutical Sciences
Oklahoma City, Oklahoma, USA
2019-07-02T04:38:05+00:00
https://www.dualsystems.com/testimonials/the-university-of-oklahoma-health-sciences-center/
CuroNZ Ltd
The team from DualSystems Biotech AG has been extremely helpful in delivering scientific advice for CuroNZ’s target identification journey. The quick turnaround time and the precision of NRP2945’s target identification in regard to the utilization of their TRICEPS technology impressed us very much. Meanwhile we could identify another NRP2945 receptor target that was identified by DualSystems as being recruited to the NRP2945-activated membrane receptor complex. An absolute pleasure to work with Paul Helbling’s team.
Frank Sieg, PhD
CSO
CuroNZ Ltd
Mangawhai in New Zealand
Frank Sieg, PhD
CSO
CuroNZ Ltd
Mangawhai in New Zealand
2019-02-20T12:18:04+00:00
https://www.dualsystems.com/testimonials/curonz-ltd/
University of Pittsburgh
It was a pleasure working with Paul Helbling and his team. I was impressed by their attention to detail, and optimization of all cell conditions to produce the most reliable possible results. They kept me informed at every stage of the process and shared data as soon as they were able to. Using their LRC-TriCEPS platform, we were able to follow leading candidates as binding partners for our cardiac targeting peptide and identified the most likely binding partner.
Maliha Zahid, M.D., Ph.D.
Assistant Professor
Departement of Developmental Biology
University of Pittsburgh
Maliha Zahid, M.D., Ph.D.
Assistant Professor
Departement of Developmental Biology
University of Pittsburgh
2018-11-15T19:35:14+00:00
https://www.dualsystems.com/testimonials/university-of-pittsburgh/
University of Oklahoma Health Sciences Center
I was very pleased with my collaboration with the team at Dualsystems. We had to work with difficult cells and the team helped our lab develop modifications to the original protocol to be able to use the TriCEPS approach with our cells. The whole team was very responsive, involved, and knowledgeable and this led to a successful identification of a receptor for our ligand of interest. We have since then confirmed this interaction and we are now getting ready to publish this study, which will have implications in the treatment of ocular injuries.
Anne Kasus-Jacobi, PhD
Assistant Professor of Research
University of Oklahoma Health Sciences Center
Department of Pharmaceutical Sciences
Oklahoma City, Oklahoma, USA
Anne Kasus-Jacobi, PhD
Assistant Professor of Research
University of Oklahoma Health Sciences Center
Department of Pharmaceutical Sciences
Oklahoma City, Oklahoma, USA
2018-04-23T11:45:16+00:00
https://www.dualsystems.com/testimonials/lrc-triceps/
Biomedical Research Institute
PUBLICATION: Li Z, Zeppa JJ, Hancock MA, McCormick JK, Doherty TM, Hendy GN, and Madrenas J. Staphylococcal Superantigens Use LAMA2 as a Coreceptor To Activate T Cells. J Immunol. 2018; 200: 1471-1479.
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.
(Quim) Madrenas, MD, PhD, FCAHS
Chief Scientific Officer
Los Angeles Biomedical Research Institute at Harbor-UCLA Medical Center
Torrance, USA
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.
(Quim) Madrenas, MD, PhD, FCAHSChief Scientific Officer
Los Angeles Biomedical Research Institute at Harbor-UCLA Medical Center
Torrance, USA
2018-04-19T11:18:18+00:00
https://www.dualsystems.com/testimonials/lrc-triceps-service-8/
QIMR Berghofer Medical Research Institute
“With the help of the team at Dualsystems we confirmed a potential receptor which has lead to a Nature paper submission and successful funding of a research grant. The LRC-TriCEPS experiment was straightforward and the process was cost effective. We look forward to using TriCEPS in the future.”
Anita Burgess | PhD
Hepatic Fibrosis Group
QIMR Berghofer Medical Research Institute, Australia
Anita Burgess | PhDHepatic Fibrosis Group
QIMR Berghofer Medical Research Institute, Australia
2017-12-12T13:21:33+00:00
https://www.dualsystems.com/testimonials/lrc-triceps-service-7/
University of Miami, Miller School of Medicine
We are very pleased with the experience and interaction on very high professional level with Dualsystems Biotech. As a result of such collaboration the new receptor MUC5B was identified in human chondrosarcoma cells for the first time for antiproliferative neuropeptide PRP-1. Read more
I would like to thank once again the company and, particularly, Dr Helbling for his attention and collaboration.
Dr Karina Galoian
Research associate professor
University of Miami, Miller School of Medicine
Department of Orthopedic surgery
Miami, Florida, USA
I would like to thank once again the company and, particularly, Dr Helbling for his attention and collaboration.
Dr Karina GaloianResearch associate professor
University of Miami, Miller School of Medicine
Department of Orthopedic surgery
Miami, Florida, USA
2017-11-16T08:07:04+00:00
https://www.dualsystems.com/testimonials/lrc-triceps-service-6/
Münster University Hospital (UKM)
"Dualsystems Biotech’s approach to receptor identification with the TriCEPS reagent gave us a powerful platform with great support for a successful and fast data generation, when conventional approaches had previously failed."
Dr. rer. nat. Martin Herold
Working group from Prof. Dr. med. Luisa Klotz
Münster University Hospital (UKM), Germany
Dr. rer. nat. Martin HeroldWorking group from Prof. Dr. med. Luisa Klotz
Münster University Hospital (UKM), Germany
2017-08-24T14:48:20+00:00
https://www.dualsystems.com/testimonials/lrc-triceps-service-5/
University of Manitoba
"Every aspect of our experience with Dualsystems Biotech was outstanding. They provided excellent customer service and technical support throughout the entire process. Their LRC-TriCEPS technology allowed us to identify several new candidate receptors for our protein of interest in rat primary neurons, and this has created new and exciting avenues for our research."
Sari S. Hannila, PhD
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
Sari S. Hannila, PhDAssociate 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
2017-06-12T10:05:43+00:00
https://www.dualsystems.com/testimonials/lrc-service/
The Rockefeller University
"We are very pleased with the work product of Dualsystems Biotech. The TriCEPS reagent allowed us to identify a novel ligand-extracellular matrix protein receptor interaction, which was not possible using traditional techniques. The Dualsystems team were very helpful in tailoring the experimental conditions to fit our biological question."
Manish Ponda, M.D., M.S.
Assistant Professor of Clinical Investigation
The Rockefeller University
Manish Ponda, M.D., M.S.Assistant Professor of Clinical Investigation
The Rockefeller University
2016-10-10T14:21:51+00:00
https://www.dualsystems.com/testimonials/lrc-triceps-service-3/
Medizinische Hochschule Hannover
“With the support of Dualsystems Biotech and their LRC-TriCEPS technology we were able to identify receptor candidates for our peptide with renoprotective properties in kidney injury.”
Dr. rer. nat. Inga Sörensen-Zender
Postdoc
Medizinische Hochschule Hannover
Dr. rer. nat. Inga Sörensen-ZenderPostdoc
Medizinische Hochschule Hannover
2016-09-26T10:26:51+00:00
https://www.dualsystems.com/testimonials/lrc-triceps-service-2/
East Tennessee State University
“For the ligand sample two unique receptors were identified. Flow cytometry analysis with siRNA induced knockdown of these proteins confirmed that the presence of the protein is needed for CTRP3 binding to occur. CONCLUSION The LRC-TriCEPS methodology was successful in identifying the receptor for CTRP3.”
Jonathan M Peterson
Assistant Professor
East Tennessee State University
Jonathan M PetersonAssistant Professor
East Tennessee State University
2016-05-30T15:20:03+00:00
https://www.dualsystems.com/testimonials/lrc-triceps-service/
Igenica Biotherapeutics
«Leveraging Dualsystems Biotech novel linker technology (LRC-TriCEPS) and its analytical data processing capabilities we were able to identify the heterophilic receptor for a highly-pursued immuno-oncology target.»
Dr. Edward van der Horst,
Senior Director, Preclinical Development
Igenica Biotherapeutics
Dr. Edward van der Horst,Senior Director, Preclinical Development
Igenica Biotherapeutics
2016-05-09T11:13:08+00:00
https://www.dualsystems.com/testimonials/lrc-triceps-service-4/
Centro de Estudos de Doenças Crónicas
New publication in Nature Communications using the LRC-TriCEPS technology in collaboration with Alisson Gontijo
« The fruitful collaboration with Dualsystems Biotech using the LRC-TriCEPS (CaptiRec) technology showed that even on insect cells receptors could be identified »
Alisson M. Gontijo,
Principal Investigator at CEDOC
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 »
Alisson M. Gontijo,Principal Investigator at CEDOC
Centro de Estudos de Doenças Crónicas
2015-02-24T07:32:40+00:00
https://www.dualsystems.com/testimonials/captirec-service-3/
Washington University School of Medicine
"Thanks to LRC-TriCEPS (CaptiRec) technology we were able to identify a long sought receptor of our ligand using genetically-engineered cell lines with the support of Dualsystems".
Fumihiko Urano, MD, PhD
Samuel E. Schechter Professor of Medicine
Washington University School of Medicine
Fumihiko Urano, MD, PhDSamuel E. Schechter Professor of Medicine
Washington University School of Medicine
2015-02-17T14:30:12+00:00
https://www.dualsystems.com/testimonials/captirec-service-2/
University of California San Francisco
"We have been trying to identify a receptor that has been sought after for nearly 30 years by laboratories across the globe. Thanks to Dualsystems service CaptiRec (LRC-TriCEPS), we now have a strong candidate receptor." 
De'Broski R. Herbert Ph.D.
Assistant Professor in Residence
University of California San Francisco (UCSF)
De'Broski R. Herbert Ph.D.Assistant Professor in Residence
University of California San Francisco (UCSF)
2015-02-17T14:27:53+00:00
https://www.dualsystems.com/testimonials/captirec-service/
LRC-TriCEPS customers worldwide
Over 200 satisfied customers from 28 countries.
