Identification of cell surface proteins (surfaceome)
by TriCEPS
TriCEPS-Surfaceome on primary human T-cells
Why is the surfaceome (sum of all proteins at the cell surface) important?
a) Cell communication: Detects and transmits cell signals
b) Cell differentiation and activation: Changes its protein expression pattern depending on internal and external signals
c) Cell protection: Keeps cell integrity and provides a boundry
d) Cell migration: Leads the way by communicating with its surrounding
e) Immunity: Key for identification of self and foreign and the diseased state of a cell
For all these functions the surface proteins (surfaceome) are key.
The surfaceome content differs among cell types and changes during developmental and disease states. Therefore, it contains unique Biomarkers that can be used to distinguish cellular phenotypes and disease states. These properties along with the fact that cell surface proteins are readily available make the surfaceome a rich source of phenotypic, diagnostic, prognostic and therapeutic targets that can be used in a variety of fields including oncology, immunology and stem cell research
Using our TriCEPS platform technology we can identify the surface proteins of any given cell and compare the quantitative changes between two cellular states.
As an example experiment, we show the quantitative changes taking place on primary human CD4+ T cells activated by CD3, CD28 and IL-2 for 3 and 11 days compared to naïve T cells.
Figure 1: As an example experiment, we show the quantitative changes taking place on primary human CD4+ T cells activated by CD3, CD28 and IL-2 for 3 days and 11 days compared to naïve T cells. The TriCEPS surfaceome workflow was conducted to identify quantitative changes of surface proteins on the naïve and activated primary human T cells.
Roughly 600 membrane associated proteins (surfaceome) were identified for the primary T cells; approximately 150 and 200 proteins showed differential expression after 3 and 11 days of activation respectively when compared to naïve T-cells.
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Volcano plots depicting the comparison between naive and activated T cells
Data is shown at the protein level and proteins were annotated using the human proteome database from Uniprot. Identified proteins were filtered and only membrane associated and/or secreted proteins were retained. Results are presented in the format of a volcano plot; Y axis = -Log10 (adj. p-value), X-axis = Log2 fold change compared to the other sample.
Proteins in the green space showed an increase in abundance upon T cell stimulation whereas proteins in the blue space were more abundant on naïve T cells.
Known T cell surface activation and proliferation markers, including receptor proteins (e.g. TFR1 and IL2RA (CD25)) co-stimulatory molecules (e.g. ICOS and TNR4 (OX40)), adhesion proteins (e.g. ICAM1) and interleukines (e.g. IL16) were found to be differentially abundant upon stimulation
Left plot: proteins known to change in abundance upon activation
Right plot: proteins constitutively expressed
Figure 2: Expression profiles of known T cell surface activation and proliferation Biomarkers across naïve and activated T cells
Customers Testimonials – LRC-TriCEPS Service
Testimonials from our customers who have used the LRC-TriCEPS technology – in collaboration with Dualsystems Biotech AG.
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.
LRC-TriCEPS / HATRIC-LRC Publications
Concerning the LRC-TriCEPS or HATRIC-LRC platforms.
