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Flow Cytometry as Platform for Biomarker Discovery and Clinical Validation

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General Methods in Biomarker Research and their Applications

Abstract

Flow cytometry (FC) is a highly versatile method that is constantly expanding. Its field of application is extremely wide (oncology, hematology, transplantation, autoimmunity, tumor immunology, chemotherapy, etc.), making it highly useful not only in the discovery of new biomarkers but also in clinical validation and routine implementation. FC simultaneously provides information about the phenotypic and functional characteristics of cells and enables quantification of large numbers of cells and assessment of their subset distribution, activation status, cytokine production profile, and other cellular functions. Furthermore, because FC allows the performance of many different types of assays (immunophenotyping, intracellular staining, cell cycle, cell proliferation, apoptosis, phosphoflow assays, etc.), it yields different types of information, ranging from elucidation of mechanisms of action (for drugs and disease progression) to functional assays. This technique plays an important role in the prioritization, verification, and clinical validation of new biomarkers. However, because of the high complexity of the panels of reagents involved, greater expertise is needed for correct interpretation of the data obtained, and the technique continues to have several limitations. One of the most important limitations is the lack of standardization in assay and instrument setup, as well as the absence of good quality controls, especially external controls. There is a need to develop mathematical algorithms able to predict clinical evolution/disease progression based on FC measurement of biomarkers. New biostatistics models should be developed to establish the most appropriate correlation among biomarkers, drug effect, and clinical outcome, which would allow personalized treatment.

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Abbreviations

AR:

Acute Rejection

BrDU:

Bromodeoxyuridine

C&T:

Cytometric Setup and Tracking

cGVHD:

Chronic Graft-Versus-Host Disease

CSC:

Cancer Stem Cell

DNA:

Deoxyribonucleic Acid

EDTA:

Ethylenediaminetetraacetic Acid

FC:

Flow Cytometry

ICS:

Intracellular Staining

IFN-γ:

Interferon-γ

IL:

Interleukin

PBMC:

Peripheral Blood Mononuclear Cells

S6RP:

S6 Ribosomal Protein

SLE:

Systemic Lupus Erythematosus

SOPs:

Standard Operating Procedures

TCR:

T-Cell Receptor

Treg:

Regulatory T Cells

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Authors and Affiliations

  1. Pharmacology and Toxicology Laboratory, Centro de Diagnóstico Biomédico, CIBERehd, IDIBAPS, Hospital Clínico de Barcelona, Barcelona University, C/ Villarroel 170, 08036, Barcelona, Spain

    Olga Millán & Mercè Brunet

Authors
  1. Olga Millán
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  2. Mercè Brunet
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Correspondence to Mercè Brunet .

Editor information

Editors and Affiliations

  1. Department of Nutrition and Dietetics, Division of Diabetes & Nutritional Sciences, Faculty of Life Sciences & Medicine, King's College London, London, United Kingdom

    Victor R. Preedy

  2. Faculty of Science & Technology, Department of Biomedical Sciences, University of Westminster, London, United Kingdom

    Vinood B. Patel

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Millán, O., Brunet, M. (2015). Flow Cytometry as Platform for Biomarker Discovery and Clinical Validation. In: Preedy, V., Patel, V. (eds) General Methods in Biomarker Research and their Applications. Biomarkers in Disease: Methods, Discoveries and Applications. Springer, Dordrecht. https://doi.org/10.1007/978-94-007-7696-8_16

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