Grupo Proteómica




Head of Unit: Gemma Barroso García



Mass spectrometry is a well-established technique and widely used in industry and academia. The technological advance of this technique has allowed obtaining a high knowledge on the characterization, function and interaction between key biomolecules for the cell such as proteins and metabolites.


The Proteomics Unit's mission is to provide researchers with state-of-the-art mass spectrometry methods to analyze possible biomolecular changes that can help them better understand diseases, drug effects, metabolic pathways, etc.
Since 2007, when Proteomics is part of the Research Support Facilities of the National Hospital of Paraplegics (SAI-HNP), new applications have been incorporated for characterization studies, massive identification, quantification or targeted studies. In addition to providing these services to the center's researchers, they are also offered to other public institutions and private companies.


The Unit participates in the continuing education program of the National Hospital of Paraplegics-SESCAM, as well as through the Foundation for the National Hospital for Paraplegics, giving theoretical-practical courses aimed at specialists, clinicians, researchers and technical personnel interested in increase their knowledge of proteomics techniques and their potential applications to their fields of interest. Likewise, there are agreements with universities to carry out internships at our facilities.


The SAI-Proteomics participates and collaborates in the organization of different informative activities developed by the Research Unit, such as SAIs Conference, Brain Week, Seminars, etc. to bring the work we develop closer to the scientific community and society.





Mass Spetrometry

TripleTOF 6600 plus (Sciex)

The TripleTOF 6600 plus is a Q-TOF mass spectrometer that provides high resolution and mass accuracy in all MSn stages. The TripleTOF is coupled with an Eksigentnano / microLC 425 system. It is suitable for independent data acquisition (DIA) and mass identification.

4000Qtrap (Sciex)

The 4000QTrap is a triple quadrupole (QqQ) mass spectrometer. These types of spectrometers are specially designed for targeted proteomics analysis (SRM / MRM). The 4000QTrap is equipped with a nano-ESI source and has a nanoLC Ultimate 3000 (Dionex) system attached.

4800 Plus MALDI TOF/TOF (Sciex)

The 4800 Plus MALDI TOF/TOF is a dual time-of-flight (TOF/TOF) analyzer mass spectrometer with a MALDI ionization source. It allows the rapid determination of molecular masses of intact peptides and proteins.

Electrophoresis and 2D-gel image analysis

Isoelectric focusing systems

The SAI has two thermostatted isoelectric focusing equipment for the separation of proteins according to their isoelectric point using immobilized pH gradient (IPG) strips of several sizes.

  • EttanIPGphor 3 (GE Healthcare)
  • PROTEAN i12 IEF Cell (BioRad)

Electrophoresis systems

The electrophoresis systems available are based on the separation of proteins through a porous matrix (gel electrophoresis) according to their molecular weight by applying them an electric field.

  • EttanDalt Six (GE Healthcare)
  • Hoefer SE 600 (GE Healthcare)
  • Mini-PROTEAN 3 Dodeca Cell and Mini-PROTEAN Tetra Cell (BioRad)

Imaging Systems

  • ImageScanner III
  • Typhoon ™ TRIO. Scanner for the visualization of fluorescent images specially designed for DIGE gels (GE Healthcare).

Other equipment

- EXQuest Spot Cutter: (BioRad).

- Äkta Prime plus (GE Healthcare).

- Software for data analysis.
The proteomics core has computing resources to process the data generated in the assays offered.

  • DeCyder ™ v7.0: specific software for differential expression analysis of proteins in gel labeled with DIGE technology (GE Healthcare).
  • ProteinPilot ™ v5.0.2 (Sciex): automated protein search program including MASCOT v2.5 (search engines and Paragon.
  • Peakview v2.0 (Sciex): software to visualize and review the data from LCMS and LCMSMS.
  • MarkerView v1.3.1 (Sciex): statistical analysis software based on mass spectrometry data.
  • Skyline(MacCoss Lab-open source): software for generating targeted proteomic methods (MRM) and subsequent analysis.







Analysis Offered

- Polyacrylamide gel electrophoresis (SDS-PAGE)

- 2D electrophoresis (2-DE)

- 2D differential quantitative electrophoresis (2D-DIGE)

- Molecular mass determination.

- Ion fragmentation analysis.

- Identification and characterization of proteins by MALDI.

- Identification and characterization of proteins by LC-MS / MS.

- Quantitative analysis:

  • SRM / MRM (Selected Reaction Monitoring / Multiple Reaction Monitoring)







- Serrano-Heras, G., Díaz-Maroto, I., Castro-Robles, B. et al. Isolation and Quantification of Blood Apoptotic Bodies, a Non-invasive Tool to Evaluate Apoptosis in Patients with Ischemic Stroke and Neurodegenerative Diseases. Biol Proced Online 22, 17 (2020).

- Martin-Rojas, T., Sastre-Oliva, T., Esclarín-Ruz, A., Gil-Dones, F., Mourino-Alvarez, L., Corbacho-Alonso, N., Moreno-Luna, R., Hernandez-Fernandez, G., Lopez, J. A., Oliviero, A., & Barderas, M. G. (2020). Effects of Growth Hormone Treatment and Rehabilitation in Incomplete Chronic Traumatic Spinal Cord Injury: Insight from Proteome Analysis. Journal of personalized medicine, 10(4), 183.

- Arevalo-Martin A, Grassner L, Garcia-Ovejero D, Paniagua-Torija B, Barroso-Garcia G, Arandilla AG, Mach O, Turrero A, Vargas E, Alcobendas M, Rosell C, Alcaraz MA, Ceruelo S, Casado R, Talavera F, Palazón R, Sanchez-Blanco N, Maier D, Esclarin A, Molina-Holgado E. Elevated Autoantibodies in Subacute Human Spinal Cord Injury Are Naturally Occurring Antibodies. Front Immunol. 2018 Oct 11;9:2365. doi: 10.3389/fimmu.2018.02365. eCollection 2018 PMID: 30364218

- Mourino-Alvarez L, Baldan-Martin M, Sastre-Oliva T, Martin-Lorenzo M, Maroto AS, Corbacho-Alonso N, Rincon R, Martin-Rojas T, Lopez-Almodovar LF, Alvarez-Llamas G, Vivanco F, Padial LR, de la Cuesta F, Barderas MG. A comprehensive study of calcific aortic stenosis: from rabbit to human samples. Dis Model Mech. 2018 Jun 19;11(6):dmm033423. doi: 10.1242/dmm.033423. PMID: 29752279; PMCID: PMC6031362.

- Martin-Rojas T, Mourino-Alvarez L, Gil-Dones F, de la Cuesta F, Rosello-Lleti E, Laborde CM, Rivera M, Lopez-Almodovar LF, Lopez JA, Akerstrom F, Padial LR, Barderas MG. A clinical perspective on the utility of alpha 1 antichymotrypsin for the early diagnosis of calcific aortic stenosis. Clin Proteomics. 2017 Apr 21;14:12.

- De la Cuesta F, Baldan-Martin M, Moreno-Luna R, Alvarez-Llamas G, Gonzalez-Calero L, Mourino-Alvarez L, Sastre-Oliva T, López JA, Vázquez J, Ruiz-Hurtado G, Segura J, Vivanco F, Ruilope LM, Barderas MG. Kalirin and CHD7: novel endothelial dysfunction indicators in circulating extracellular vesicles from hypertensive patients with albuminuria. Oncotarget. 2017 Feb 28;8(9):15553-15562.

- Cantillo J, F, Puerta L, Lafosse-Marin S, Subiza J, L, Caraballo L, Fernández-Caldas E: Identification and Characterization of IgE-Binding Tropomyosins in Aedes aegypti. Int Arch Allergy Immunol 2016;170:46-56. doi: 10.1159/000447298

- Baldan-Martin M, Mourino-Alvarez L, Gonzalez-Calero L, Moreno-Luna R, Sastre-Oliva T, Ruiz-Hurtado G, Segura J, Lopez JA, Vazquez J, Vivanco F, Alvarez-Llamas G, Ruilope LM, de la Cuesta F, Barderas MG. Plasma Molecular Signatures in Hypertensive Patients With Renin-Angiotensin System Suppression: New Predictors of Renal Damage and De Novo Albuminuria Indicators. Hypertension. 2016 Jul;68(1):157-66. doi: 10.1161/HYPERTENSIONAHA.116.07412. Epub 2016 May 23. PMID: 27217411.

- Pita-Thomas W, Barroso-García G, Moral V, Hackett AR, Cavalli V, Nieto-Diaz M. Identification of axon growth promoters in the secretome of the deer antler velvet Neuroscience. 2016 Nov 4. pii: S0306-4522(16)30604-2 Citada en PubMed; PMID: 27818162

- Sánchez-Sierra M, García-Álvarez I, Fernández-Mayoralas A, Moreno-Lillo S, Barroso García G, Moral Dardé V, Doncel-Pérez E. Mass Spectrometry in Pharmacokinetic Studies of a Synthetic Compound for Spinal Cord Injury Treatment. Biomed Res Int. 2015;2015:169234 Citada en PubMed; PMID: 26090386

- REGIONAL HETEROGENEITY OF 2-AG SYNTHESIZING AND DEGRADING MACHINERIES IN OLIGODENDROCYTES. INTRODUCING MULTIPLE REACTION MONITORING FOR QUANTIFICATION OF PROTEINS OF THE ENDOCANNABINOID SYSTEM Esteban PF:, Garcia-Ovejero D., Moreno-Luna R., Paniagua-Torija B., Arandilla AG., Barroso-García G., Arévalo-Martín A., Molina-Holgado E., 19a Reunión Anual de la Sociedad Española de Investigación sobre Cannabinoides, 22-24 Nov 2018 Madrid

- AUTOANTIBODIES AGAINST CENTRAL NERVOUS SYSTEM AFTER SPINAL CORD INJURY Angel Arevalo-Martin, Daniel Garcia-Ovejero, Beatriz Paniagua-Torija, Lukas Grassner, Orpheus Mach, Eduardo Vargas Baquero, Monica Alcobendas Maestro, Ana Esclarín de Ruz, Gemma Barroso García, Eduardo Molina-Holgado, 40 Congreso de la Sociedad Española de Inmunología, 25-27 Mayo 2017 Zaragoza

- AUTOANTIBODY SIGNATURES RELATED WITH RECOVERY AFTER SPINAL CORD INJURY Angel Arevalo-Martin, Lukas Grassner, Daniel Garcia-Ovejero, Beatriz Paniagua-Torija, Orpheus Mach, Gemma Barroso García, Eduardo Vargas Baquero, Monica Alcobendas Maestro, Dorsi Maier, Eduardo Molina-Holgado Cell Symposia. Neuro-Inmune Axis: Reciprocal Regulation in Development, Health, and Diseases 17-19 Sept. 2017 Sitge

- SPINAL CORD INJURY INDUCES LONG-TERM CHANGES IN THE RAT SPLEEN PROTEOME Angel Arevalo-Martin, Daniel Garcia-Ovejero, Veronica M Darde, Gemma Barroso, Yolanda Sierra-Palomares, Eduardo Molina-Holgado. 3rd International Spinal Cord Repair Meeting,  6-7 noviembre 2015 Barcelona.





Barroso García, Gemma. Head of Unit



González Arandilla, Alba. Senior Mass Spectrometry



Gallardo Escobar, Ana. Sample Preparation






Proteomics Unit

Edf. Investigación i-1Lab 21

National Paraplegic Hospital

Finca La Peraleda s/n

45071 Toledo




Tfno: (+34) 925 396 823

Tfno 2: (+34) 638 228 821

Fax: (+34) 925 396 821