The Immune Response and Biomarkers Facility provides advanced quantification of a wide range of biomarkers across diverse biospecimens, using technologies such as ELISA, xMAP Luminex and targeted proteomics with OLINK (Certified International Core Lab).

The biomarkers analysed are primarily proteins, including antibodies of different isotypes and subclasses against customised antigen panels, as well as other immune-related analytes such as cytokines, chemokines and growth factors. In addition, biomarkers associated with other physiological pathways, including metabolic, neurological and cardiovascular processes, can also be assessed.

Key strengths of the facility include:

• Multiplexing capacity, enabling multiparametric data generation from single samples (Luminex and OLINK).
• Miniaturisation, suitable for studies with limited sample volumes (e.g. paediatric cohorts).
• Versatility and adaptability (customized needs, kits can be commercial or produced in-house).
• High-throughput capacity through semiautomated liquid handling systems.
• Functional antibody assays (e.g. binding inhibition and surrogate neutralisation).
• Complementarity of technologies, allowing cross-validation of biomarkers (e.g. Luminex vs OLINK for cytokines and other biomarkers).

Custom panels are developed using recombinant proteins (commercial or produced in-house in prokaryotic or eukaryotic systems) and monoclonal antibodies (mAbs) for customised applications. These enable in-depth immune phenotyping, using spectral cytometry, and the analysis of T-cell specific responses through intracellular cytokine staining (ICS) and T-cell activation-induced marker (AIM) assays by flow cytometry and ELIspot/FluoroSpot assays.

The following specific knowledge and capacities are offered as added values:

• Extensive expertise and experience in immune monitoring.
• A long track record of the team.
• Built-in QA/QC and data preprocessing R pipelines.
• The possibility to perform statistical analyses, consulting, training, and capacity building for 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).
• Other cellular assays (T cells, B cells) upon request (flow cytometry, ELIspot/FluoroSpot) For these technologies we offer:
  • Advice on what markers, technologies (ELISA, Luminex, Olink) and kits to use depending on the objective and specificities of the study.
  • Design of experiments.
  • Supplier contacts and preparation of quotations.
  • Purchase of reagents and kits.
  • Performance of the assays.
  • QA/QC of the data generated.
  • Additionally: Flow cytometry and ELIspot/FluoroSpot (sample acquisition/reading using IDIBAPS platform instruments).

Equipment

• CytoFLEX (Beckman Coulter)
• Access to the PCB flow cytometry unit (Cytek Aurora; 4 lasers, spectral)
• Access to the PCB ELISA microplate reader (TECAN Infinite® M Plex microplate)
• Two automatic washers (BioTek)
• Luminex instruments: one Magpix, one Luminex 100/200 and two FlexMap3D
• Accessories for the Luminex assays:
  • Kingfi sher Flex 96 deepwell (Thermo Fisher) for automated antigen coupling
  • Integra Assist Plus and Integra Viafl o 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)
• 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)
• Softwares for running ELISA (GraphPad Prism), Luminex (X-ponent 4.3), Olink (NPX Signature software), and fl ow cytometry (FlowJo)
• Ella™ automated ELISA (Bio-Techne)

This core facility is part of the CERCAGINYS platform.

Quality Control

• Quality Policy
• Certificate ISO 9001
• Certificate Olink

Selected Projects

• ATHLETE - Advancing Tools for Human Early Lifecourse Exposome Research and Translation
  Funding organisation: EU-H2020
  Status: Executing
• BISC - Barcelona Life Study Cohort
  Funding organisation: EIC-HEI
  Status: Finished
• COVICAT - Cohort Covid-19 a Catalunya
  Funding organisation: CERCA, EIT Health, “la Caixa” Social Research Grant SR20-01024
  Status: Executing
• EPHOR - Exposome Project for Health and Occupational Research
  Funding organisation: EU-TNO (NED)
  Status: Executing
• BASEMALVAC - Baseline host and environmental factors that impact pre-erythrocytic malaria vaccine (hypo)responsiveness in endemic regions
  Funding organisation: NIAID-NIH (USA) et al.
  Status: Executing
• END-VOC - Ending COVID-19 variants of concern through cohort studies
  Reference: HORIZON-HLTH-2021-CORONA-01-02
  Funding organisation: EU-HE
  Status: Executing
• INCENTIVE - Indo-European Consortium for Next Generation Influenza Vaccine Innovation
  Funding organisation: EU-H2020
  Status: Executing
• 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 organisation: ISCIII
  Status: Finished
• Infection acquisition in early life and health outcomes in childhood
  Funding organisation: Fundació La Marató de TV3
  Status: Finished
• CoViproteHCt - Identifying protective immunity to SARS-CoV-2 in Health Care Personnel
  Funding organisation: EIT Health
  Status: Finished
• 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 organisation: UNITAID
  Status: Finished

Selected Publications

• Evaluation of the accuracy of a multi-infection screening test based on a multiplex immunoassay targeting imported diseases common in migrant populations
  Aguilar R, Cruz A, Jiménez A, et al.; Requena-Méndez A.
  Travel Medicine and Infectious Disease. 2023;57:102681.
  DOI: 10.1016/j.tmaid.2023.102681
• Individual blood concentrations of persistent organic pollutants and chemical elements, and COVID-19: A prospective cohort study in Barcelona
  Porta M, Pumarega J, Gasul M, et al.; Moncunill G, Aguilar R.
  Environmental Research. 2023;223.
  DOI: 10.1016/j.envres.2023.115419
• Long-term exposure to air pollution and COVID-19 vaccine antibody response in a general population cohort (COVICAT Study, Catalonia)
  Kogevina M, Karachaliou M, Espinosa A, et al.; Dobaño C.
  Environmental Health Perspectives. 2023;131(4):04700.
  DOI: 10.1289/EHP11989
• RBD-based ELISA and Luminex predict anti-SARS-CoV-2 surrogate-neutralising activity in two longitudinal cohorts of German and Spanish healthcare workers
  Aguilar R, Li X, Crowell CS, et al.; Gerhard M.
  Microbiology Spectrum. 2023;11(1):e0316522.
  DOI: 10.1128/spectrum.03165-22
• Evaluation of antibody serology to determine current helminth and Plasmodium falciparum infections in a co-endemic area in southern Mozambique
  Santano R, Rubio R, Grau-Pujol B, Moncunill G.
  PLoS Neglected Tropical Diseases. 2023;16(6):e0010138.
  DOI: 10.1371/journal.pntd.0010138
• SARS-CoV-2 infection, vaccination, and antibody response trajectories in adults: A cohort study in Catalonia
  Karachaliou M, Moncunill G, Espinosa A, Dobaño C.
  BMC Medicine. 2022;20(1):347.
  DOI: 10.1186/s12916-022-02547-2
• HIV infection and placental malaria reduce maternal transfer of multiple antimalarial antibodies in Mozambican women
  Alonso S, Vidal M, Ruiz-Olalla G, Moncunill G.
  Journal of Infection. 2021;82(4):45–57.
  DOI: 10.1016/j.jinf.2021.02.024
• Highly sensitive and specific multiplex antibody assays to quantify immunoglobulins M, A, and G against SARS-CoV-2 antigens
  Dobaño C, Vidal M, Santano R, Moncunill G.
  Journal of Clinical Microbiology. 2021;59(2).
  DOI: 10.1128/JCM.01731-20
• Seven-month kinetics of SARS-CoV-2 antibodies and role of pre-existing antibodies to human coronaviruses
  Ortega N, Ribes M, Vidal M, Dobaño C.
  Nature Communications. 2021;12(1):4740.
  DOI: 10.1038/s41467-021-24979-9
• Antibody responses to the RTS,S/AS01E vaccine and Plasmodium falciparum antigens after a booster dose within the phase 3 trial in Mozambique
  Sánchez L, Vidal M, Jaroice C, Dobaño C.
  NPJ Vaccines. 2020;5:46.
  DOI: 10.1038/s41541-020-0192-7
• A balanced proinflammatory and regulatory cytokine signature in young African children is associated with lower risk of clinical malaria
  Dobaño C, Nhabomba AJ, Manaca MN, Alonso PL.
  Clinical Infectious Diseases. 2019;69(5):820–828.
  DOI: 10.1093/cid/ciy934