Immune Response and Biomarkers
GINYS-ISGLOBAL-004
Carrer Roselló 153 (CEK building, 1st floor)
E-08036 Barcelona, Catalonia, Spain
E-08036 Barcelona, Catalonia, Spain
This Core Facility offers as service the quantification of a large panel of biomarkers through different technologies like ELISA and x-MAP Luminex, as well as targeted proteomics with OLINK (Certified International Core lab), in diverse biospecimens. The biomarkers profiled are mainly proteins, including antibodies of different isotypes and subclasses against customized panels of antigens, and other immune analytes like cytokines, chemokines, and growth factors. Biomarkers involved in other physiological pathways like metabolic, neurologic, cardiovascular, etc, processes are also measured.
The highlights of the core facility include: i) the multiplexing capacity (multiparametric information from single samples in Luminex and Olink), ii) miniaturization suitable for small sample volumes (e.g. pediatric cohort studies), iii) the versatility and adaptability (customized needs, kits can be commercial or produced in-house), iv) the high-throughput (semi-automated liquid handling equipment), v) the functional antibody capacity (e.g. inhibition of binding, surrogate neutralization assays), and vi) the complementarity of techniques (e.g. Luminex vs. Olink for cytokines and other biomarkers).
In our panels we use commercial recombinant proteins or produced in-house in prokaryotic or eukaryotic expression systems, and monoclonal antibodies (mAbs), for customized applications, to perform in depth cell immune phenotyping using spectral cytometry, and to analyse T cell specific responses using intracellular cytokine staining (ICS) and T-cell activation-induced marker (AIM) assays by flow cytometry and ELIspot/FluoroSpot assays.
Specific knowledge and capacities offered as added values include extensive expertise and experience in immune monitoring, long track record of the team, the QA/QC and data preprocessing R pipelines built-in, and the possibility to perform statistical analyses, consulting, training and capacity building, including LMIC laboratory personnel.
Services
– ELISA and colorimetric assays using commercial kits or developed in-house to measure concentrations of different analytes.
– Multiplex cytokine Luminex assays using commercial kits to measure concentrations of different proteins like cytokines, chemokines, growth factors and other, or gene expression by Invitrogen™ QuantiGene™ Plex panels.
– Multiplex antibody Luminex assays developed by our group to measure levels and avidity of IgM, IgA, IgA1, IgA2, IgG, IgG1, IgG2, IgG3, IgG4, IgE against flexible panels of antigens from different microbes.
– Multiplex functional antibody Luminex assays.
– Olink assays using commercial kits to measure levels of up to 92 biomarkers in multiplex format (T96, T48 and 21-Flex) (Certified International CoreLab: https://olink.com/products-services/sample-analysis-service/external-service-providers/).
– Other cellular assays (T cells, B cells) upon request (flow cytometry, ELIspot/FluoroSpot)
For these technologies we offer:
a) Advice on what markers, technologies (ELISA, Luminex, Olink) and kits to use depending on the objective and specificities of the study
b) Design of experiments
c) Supplier contacts and preparation of quotations
d) Purchase of reagents and kits
e) Performance of the assays
f) QA/QC of the data generated
g) Data pre-processing
h) Preparation of the report of results. Option of statistical analysis of the data
i) Advice on interpretation of results
j) Additionally: Training and capacity building in pre-processing and analysis of data, including LMIC for in house Luminex and cellular assays.
k) Additionally: Expression of proteins and mAbs to provide to customers or for antigen-specific antibody assays (customized).
m) Additionally: Flow cytometry and ELIspot/FluoroSpot (sample acquisition/reading using IDIBAPS platform instruments).
Equipment
– ELISA microplate reader (EPOCH2, BioTek)
– Two automatic washers (BioTek)
– Luminex instruments: one Magpix, one Luminex 100/200 and two FlexMap3D – Accessories for the Luminex assays:
- Kingfisher Flex 96 deepwell (Thermo Fisher) for automated antigen coupling
- Integra Assist Plus and Integra Viaflo 384 (Integra) for liquid handling in 384 well plates
- Two 96 well plate orbital shakers (Thermo Fisher)
- Two 384 well plate orbital shakers (Heidolph Titramax: https://www.milian.com/product/0O-43-20?language=en)
– Guava easyCyte for automatic bead and cell counting
– OLINK Signature Q100
– Accessories for the OLINK assays:
- Signature Q100 Interface plate: 96.96, 48.48 and 24.192
- Centrifuge for plates (Eppendorf (5430/5430R)
- Microcentrifuge for strips and tubes (IKA Mini G Centrifuge, Fisher Scientific)
- Controlled plate vortex (MaxMate Eppendorf 230V Version)
- Tube holder for controlled plate vortex (MixMate PCR 96 tube holder, Eppendorf)
- Single and multichannel precision pipettes (Rainin, Mettler Toledo)
- Freezing block (-20ºC) for enzyme handling
- PCR instrument (SimpliAMP Thermal cycler, Applied Biosystems)
– Access to IDIBAPS cytometry platform (FACS FORTESSA 4 and 5 lasters, Aurore Cytek Cytometry)
– Softwares for running ELISA (GraphPad Prism), Luminex (X-ponent 4.3), Olink (NPX Signature software: https://olink.com/products-services/data-analysis-products/olink-npx-signature/), and flow cytometry (FlowJo)
Staff
Ruth Aguilar | ruth.aguilar@isglobal.org
| ORCID
Marta Vidal | marta.vidal@isglobal.org
| ORCID
Felipe Raimondi | felipe.raimondi@isglobal.org
Cèlia Torres | celia.torres@isglobal.org
Projects
Title:
ATHLETE - Advancing Tools for Human Early Lifecourse Exposome Research and Translation
Reference:
https://www.isglobal.org/ca/-/athlete
Funding Organism:
EU-H2020
Status:
Executing
Title:
BISC - Barcelona Life Study Cohort
Reference:
http://www.biscproject.org
Funding Organism:
EIC-HEI
Status:
Finished
Title:
COVICAT - Cohorte Covid-19 en Cataluña
Reference:
https://www.isglobal.org/ca/-/covicat
Funding Organism:
CERCA, EIT-Health,”la Caixa” Social Research grant SR20-01024
Status:
Executing
Title:
EPHOR - Exposome Project for Health and Occupational Research
Reference:
https://www.ephor-project.eu/
Funding Organism:
EU-TNO (NED)
Status:
Executing
Title:
EChiLiBRiST - Development and validation of a quantitative point-of-care test for the measurement of severity biomarkers to improve risk stratification of fever syndromes and enhance child survival
Funding Organism:
EU-HE
Status:
Executing
Title:
Baseline host and environmental factors that impact pre-erythrocytic malaria vaccine (hypo)responsiveness in endemic regions
Funding Organism:
NIAID-NIH( USA) et al.
Status:
Executing
Title:
END-VOC - Ending COVID-19 variants of concern through cohort studies
Reference:
HORIZON-HLTH-2021-CORONA-01-02
Funding Organism:
EU-HE
Status:
Executing
Title:
INCENTIVE - Indo-European Consortium for next generation influenza vaccine innovation
Funding Organism:
EU-H2020
Status:
Executing
Title:
Impact of coinfections on the balance of antibody and T helper responses to antigen targets of naturally-acquired and vaccine immunity to major human pathogens
Funding Organism:
ISCIII
Status:
Finished
Title:
Infection acquisition in early life and health outcomes in childhood
Reference:
https://www.isglobal.org/ca/-/subclinical-infections-in-children-and-long-term-health-effects
Funding Organism:
Maratò TV3
Status:
Finished
Title:
CoViproteHCt - Identifying protective immunity to SARS-CoV-2 in Health Care Personnel
Funding Organism:
EIT Health
Status:
Finished
Title:
ANTICOV-IMMUNO -The impact of COVID-19 treatment on the type, strength and duration of antibody and cellular immune responses in SARS-CoV-2 patients in sub-Saharan Africa
Funding Organism:
UNITAID
Status:
Finished
Publications
Evaluation of the accuracy of a multi-infection screening test based on a multiplex immunoassay targeting imported diseases common in migrant populations.
Travel Med Infect Dis. 2023 Dec 21;57:102681.
Travel Med Infect Dis. 2023 Dec 21;57:102681.
Individual blood concentrations of persistent organic pollutants and chemical elements, and COVID-19: A prospective cohort study in Barcelona.
Enviromental Research 2023; 223
Enviromental Research 2023; 223
Long-Term Exposure to Air Pollution and COVID-19 Vaccine Antibody Response in a General Population Cohort (COVICAT Study, Catalonia).
Environ Health Perspectives. 2023; 131-4, pp.4700
Environ Health Perspectives. 2023; 131-4, pp.4700
RBD-Based ELISA and Luminex Predict Anti-SARS-CoV-2 Surrogate-Neutralizing Activity in Two Longitudinal Cohorts of German and Spanish Health Care Workers.
Microbiol Spectrum. 2023; 14-11(1), pp.e0316522.
Microbiol Spectrum. 2023; 14-11(1), pp.e0316522.
Evaluation of antibody serology to determine current helminth and Plasmodium falciparum infections in a co-endemic area in Southern Mozambique.
PLoS Neglected Tropical Diseases. 2023; 16-6, pp.e0010138.
PLoS Neglected Tropical Diseases. 2023; 16-6, pp.e0010138.
SARS-CoV-2 infection, vaccination, and antibody response trajectories in adults: a cohort study in Catalonia.
BMC Medicine. 2022; 20-1, pp.347.
BMC Medicine. 2022; 20-1, pp.347.
Antibody conversion rates to SARS-CoV-2 in saliva from children attending summer schools in Barcelona, Spain.
BMC Medicine. 2021; 19-1, pp.309.
BMC Medicine. 2021; 19-1, pp.309.
HIV infection and placental malaria reduce maternal transfer of multiple antimalarial antibodies in Mozambican women.
Journal of Infection. 2021; 82-4, pp.45-57.
Journal of Infection. 2021; 82-4, pp.45-57.
Highly Sensitive and Specific Multiplex Antibody Assays To Quantify Immunoglobulins M, A, and G against SARS-CoV-2 Antigens.
Journal of Clinical Microbiology. 2021; 59-2.
Journal of Clinical Microbiology. 2021; 59-2.
Seven-month kinetics of SARS-CoV-2 antibodies and role of pre-existing antibodies to human coronaviruses.
Nature Communications. 2021; 12-1, pp.4740.
Nature Communications. 2021; 12-1, pp.4740.
Antibody responses to the RTS,S/AS01E vaccine and Plasmodium falciparum antigens after a booster dose within the phase 3 trial in Mozambique.
NPJ Vaccines. 2020; 5, pp.46.
NPJ Vaccines. 2020; 5, pp.46.
A Balanced Proinflammatory and Regulatory Cytokine Signature in Young African Children Is Associated With Lower Risk of Clinical Malaria.
Clinical Infectious Diseases. 2019; 69-5, pp.820-828.
Clinical Infectious Diseases. 2019; 69-5, pp.820-828.
