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Preclinical validation tissue, biomaterialls and surfaces characterization -Services

Preclinical validation tissue, biomaterialls and surfaces characterization -Services

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U30-S08. Luminex or ELISA assay development and analysis

Luminex or ELISA assay development and analysis

Design and development of immunoassays employing antibody pairs for the detection and absolute quantification of target proteins. These can be set up on a traditional ELISA format, or coupled to magnetic beads (xMAP, Luminex) for multiplex detection. Optimization procedures, including cross-reactivity assays, are performed to standardize the assay protocol. Alternatively, this service may also use commercial ELISA or multiplex kits for the analysis of specific protein panels.

Customer benefits

This service provides quantitative data of proteins and small protein panels, including cytokines, chemokines or specific biomarker candidates, in biological samples. The assays are run with high sensitivity and throughput.

Target customer

Organizations and research groups that aim to quantify in an absolute way single proteins or small panels in large number of samples. Groups interested in cytokine or chemokine profiling.

U30-S08 : Luminex or ELISA assay development and analysis
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U30-S07. Protein separation by one- or two-dimensional gel electrophoresis

Protein separation by one- or two-dimensional gel electrophoresis

Separation of complex protein mixtures on isoelectric focusing strips (according to their charge) and/or SDS-PAGE gels (according to their size), as a step for further protein or proteomic analysis.

Customer benefits

This service allows the fractionation of complex protein samples prior to proteomic analysis, performing gel-based quantitative proteomic studies by DIGE (differential in-gel electrophoresis), and evaluating protein profiles by SDS-PAGE followed by gel staining.

Target customer

Organizations and research groups interested in protein or proteomic analysis of biological samples.

U30-S07 Protein separation by one- or two-dimensional gel electrophoresis
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U30-S06. Protein or peptide separation by liquid chromatography

Protein or peptide separation by liquid chromatography

Separation of complex protein or peptide mixtures using chromatography columns. The chromatographic services offered by the Unit include strong cation exchange (IEX), reversed phase (C18), gel filtration and immunoaffinity for the removal of the top 14 most abundant proteins in human plasma.

Customer benefits

Fractionation of complex protein or peptide samples prior to proteomic or protein analysis. Processing of serum, plasma and other biofluids for shotgun proteomics analysis through the depletion of their most abundant proteins.

Target customer

This strategy is employed to verify or validate biological hypotheses with the highest specificity, as it detects the protein sequence and is not dependent on antibodies.

U30 S06 Protein or peptide separation by liquid chromatography
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U30-S05. Targeted proteomics analysis by MRM or PRM-MS

Targeted proteomics analysis by MRM or PRM-MS

Relative or absolute quantification of individual proteins or protein panels by the analysis of their peptides together with stable isotope labelled (SIL) standards.

Customer benefits

This strategy is employed to verify or validate biological hypotheses with the highest specificity, as it detects the protein sequence and is not dependent on antibodies. The use of isotope labelled standards allows the absolute quantification of the protein/s of interest.

Target customer

Organizations and research groups interested in the analysis and quantification of a protein or panel of proteins with high specificity.

U30-S05 Targeted proteomics analysis by MRM or PRM-MS
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U30-S04. Quantitative shotgun proteomics by nLC-MS/MS

Quantitative shotgun proteomics by nLC-MS/MS

Large-scale relative quantification of proteins in biological samples by nano-flow high performance liquid chromatography combined with mass spectrometry. It can use either data-dependent (DDA) or data-independent (DIA) acquisition modes. Quantifications can be performed via label-free (LFQ) or protein/peptide isotopic labelling using SILAC, iTRAQ or TMT reagents.

Customer benefits

The output of this analyses are lists of proteins identified in the samples, together with quantitative information obtained from the area displayed by each peptide in the mass spectra, and can then be compared across experimental conditions.

Target customer

Organizations and research groups interested in biomarker discovery, drug discovery (protein target identification), development of precision medicine approaches, etc.

U30-S04 Quantitative shotgun proteomics by nLC-MS/MS
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U30-S03. Protein identification by mass spectrometry (MS) or MS/MS

Protein identification by mass spectrometry (MS) or MS/MS

Identification of proteins in biological samples, either by intact protein analysis (MALDI-TOF) or protein digestion followed by peptide mass fingerprinting (PMF, MS) or peptide fragmentation (MS/MS).

Customer benefits

Identification of proteins by mass spectrometry allows the characterization of biological samples, analysis of protein interactions (e.g. pull-down assays), peptide sequencing, etc. Identifications are performed with the highest specificity.

Target customer

Organizations and research groups interested in small or large-scale protein analysis.

Additional information

E03 MALDI-TOF/TOF System

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U30-S02. Determination of molecular mass of peptides, proteins and other biomolecules

Determination of molecular mass of peptides, proteins and other biomolecules

Analysis of intact proteins, peptides and other biomolecules such as oligonucleotides using mass spectrometry (MALDI-TOF or ESI-Q-TOF) for the determination of their exact molecular mass.

Customer benefits

This service is useful to characterize a variety of biomolecules by the determination of their molecular mass with an extreme high precision. This is an essential step in many biotechnological strategies involving protein synthesis and production.

Target customer

Organizations or research groups aiming to characterize specific proteins, peptides or other biomolecules by the determination of their molecular mass.

U30 S02 Determination of molecular mass of peptides, proteins and other biomolecules
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U30-S01. Sample preparation for proteomic analysis

Sample preparation for proteomic analysis

This service includes all necessary steps to prepare samples for different proteomic analyses: protein extraction from tissues, cells or fluids, precipitation, cleaning, desalting, quantification and digestion (in gel or in solution) with proteolytic enzimes (trypsin, chymotrypsin, lys-C…), to obtain the correspondent peptides.

Customer benefits

Biological samples are processed and prepared following standardized protocols by personnel highly specialized in techniques for protein analysis.
The service allows organizations without prior experience in protein or proteomic analysis to avoid performing any experiment, only needing to provide the biological specimen to the Unit.

Target customer

Organizations or research groups interested in performing protein or proteomic analyses in a wide range of biological samples: cells, tissues, biofluids, protein extracts, etc.

U30 S01 Sample preparation for proteomic analysis
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U19-S03. Clinical Analysis Laboratory

Clinical Analysis Laboratory

The clinical analysis laboratory that performs a wide variety of analysis: (biochemistry, hematology, coagulation, urine analysis, blood gas analysis, different biomarkers, hormones, allergens) in various animal species (rat, mouse, rabbit, pig, sheep, dog, and cat). Specific methodologies are used in each animal species, having all techniques validated according to their physiological and pathological identity. It also has a formulation laboratory equipped to formulate, reconstitute and guarantee the correct storage of the test products used in these studies.

Customer benefits

The laboratories are certified with ISO-9001 and Good Laboratory Practices (GLP), strict quality standards that allow the production of high-precision results.
Also, the studies can be performed under regulatory requirements too, since the performing institution is Certified for Good Laboratory Practices.
Therefore, studies can be carried out in compliance with the strict guidelines of regulatory agencies, ensuring the reliability and traceability of all results and tests carried out in their different services.

Target customer

The services offered in this unit may be of interest to different companies and laboratories that work within the pharmaceutical industry. Companies whose objective is to test possible candidates for molecules, drugs or medical devices.
With this service, companies and laboratories ensure that they report reliable and reproducible results in all their studies, guaranteeing the traceability and viability of analyzes obtained.

References

  1. J Bote, et al. Novel ex-vivo database of a murine model of colorectal cancer using optical coherence tomography. Surg Endosc (2022) 36:S325–S674
  2. V Lucas-Cava, et al. Prostatic artery occlusion: Initial findings on pathophysiological response in a canine prostate model. Translational Andrology and Urology. Transl Androl Urol 2022.
  3. Soria F, et al. Heparin coating in biodegradable ureteral stents does not decrease bacterial colonization-assessment in ureteral stricture endourological treatment in animal model. Transl Androl Urol. 2021 Apr;10(4):1700-1710.
  4. Baez-Díaz C, et al. Microencapsulated Insulin-Like Growth Factor-1 therapy improves cardiac function and reduces fibrosis in a porcine acute myocardial infarction model. Sci Rep. 2020 Apr 28;10(1):7166.
  5. Moreno-Lobato, B, et al. Use of nanomedicine in preclinical wound healing studies. FarmaJournal; Salamanca T 4, N.º 1, (Feb 2019): 178.
  6. Picado Román, N, et al. Drugs most used in experimental animals in a Research Center. FarmaJournal; Salamanca T 4, N.º 1, (Feb 2019): 236.
  7. Moreno-Lobato B, et al. Analytical Validation Study of Hematological Parameters under Good Laboratory Practice Regulations in Different Laboratory Animal Species. Thromb Haemost Res. 2019; 3(1): 1018.
  8. Ballestín A, et al. A Pre-clinical Rat Model for the Study of Ischemia reperfusion Injury in Reconstructive Microsurgery. J Vis Exp. 2019 Nov 8;(153).
  9. R Blázquez , et al. Altered hematological, biochemical and immunological parameters in a porcine myocardial infarction model: predictive biomarkers for the severity of myocardial infarction. Veterinary Immunology and Immunopathology 205 (2018) 49–57.
  10. Vela FJ, et al. Evaluation of antigen-induced synovitis in a porcine model: Immunological, arthroscopic and kinetic studies. BMC Vet Res. 2017. Apr 7;13(1):93.
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U14-S06. Lymphocyte proliferation assay

Lymphocyte proliferation assay

The lymphocyte proliferation assay service offers a robust platform for evaluating the proliferative responses of lymphocytes in vitro. Utilizing state-of-the-art stimulation protocols and specialized proliferation assays, this service enables precise quantification of lymphocyte activation and proliferation rates. By mimicking physiological conditions in a controlled laboratory setting, researchers can investigate the dynamic behavior of lymphocytes in response to various stimuli. This service plays a pivotal role in elucidating immune responses, assessing immune competence, and evaluating the efficacy of immunomodulatory interventions, ultimately contributing to advancements in both basic immunology and clinical therapeutics.

Customer benefits

U14 Cell Therapy Unit offers researchers precision, efficiency, and expertise, streamlining experiment. With stringent quality control, ISO 9001-2015 and GLP compliant, tailored solutions, and access to advanced equipment, researchers can focus on innovation and core objectives, driving scientific progress forward.

Target customer

This service is essential for organizations involved in Research and Development, including pharmaceutical companies, academic institutions for biomedical research, biotechnology firms for bioprocessing, and regulatory agencies for in vitro studies.

Additional information

Selected references:

  • de Pedro, Maria Ageles; Pulido, Maria; Álvarez, Veronica; Marinaro, Federica; Marchena, Ana Maria; Sanchez-Margallo, Francisco Miguel; Casado, Javier G.; Lopez, Esther.  2023.  Menstrual blood-derived stromal cells: insights into their secretome in acute hypoxia conditions. MOLECULAR MEDICINE. SPRINGER. 29-1. ISSN 1076-1551.  https://doi.org/10.1186/s10020-023-00646-1
  • Cristobal JI; Duque, FJ; Uson-Casaus J; Barrera R; López E; Perez-Merino EM.  2022.  Complete Blood Count-Derived Inflammatory Markers Changes in Dogs with Chronic Inflammatory Enteropathy Treated with Adipose-Derived Mesenchymal Stem Cells   Animals.  MDPI.  12-20.  ISSN 2076-2615.  https://doi.org/10.3390/ani12202798
  • Marinaro, Federica; Silva, Joana M; Barros, Alexandre A; et al; Lopez, Esther.  2021.  A Fibrin Coating Method of Polypropylene Meshes Enables the Adhesion of Menstrual Blood-Derived Mesenchymal Stromal Cells: A New Delivery Strategy for Stem Cell-Based Therapies.International journal of molecular sciences. MDPI. 22-24. ISSN 1422-0067. WOS (0) https://doi.org/10.3390/ijms222413385
  • de Pedro, Maria Angeles; Gomez-Serrano, Maria; Marinaro, Federica; et al; Lopez, Esther (AC); Casado, Javier G.(4/10).  2021.  IFN-Gamma and TNF-Alpha as a Priming Strategy to Enhance the Immunomodulatory Capacity of Secretomes from Menstrual Blood-Derived Stromal Cells   INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES.  MDPI.  22-22.  ISSN 1422-0067.  WOS (0) https://doi.org/10.3390/ijms222212177
  • López E; Blazquez R; Marinaro F; et al; Casado JG. 2019. The Intrapericardial Delivery of Extracellular Vesicles from Cardiosphere-Derived Cells Stimulates M2 Polarization during the Acute Phase of Porcine Myocardial Infarction. STEM CELL REVIEWS AND REPORTS. SPRINGER. 16-3, pp.626-626. ISSN 2629-3269.  https://doi.org/10.1007/s12015-019-09926-y

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