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.
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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
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Left plot: proteins known to change in abundance upon activation
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Right plot: proteins constitutively expressed
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Figure 2: Expression profiles of known T cell surface activation and proliferation Biomarkers across naïve and activated T cells
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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.
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
Dr Patric Delhanty
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.(Quim) Madrenas, MD, PhD, FCAHS
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.
Dr Karina Galoian
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
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
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 »
Alisson M. Gontijo,
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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Concerning the LRC-TriCEPS or HATRIC-LRC platforms.