Name of the laboratory
- CNB-CSIC Protein Tools Unit
- Centro Nacional de Biotecnología (CNB)
- Spanish National Research Council (CSIC)
- Darwin 3, Campus Cantoblanco
- 28049 Madrid, Spain
- (+34) 915 854 614
Members of the laboratory
- Head of the Unit
Leonor Kremer (PhD) firstname.lastname@example.org
Mónica García-Gallo (PhD) email@example.com
María Teresa Martín (PhD) firstname.lastname@example.org
Ana María García Cabrero (PhD) email@example.com
Mercedes Llorente (BSc) firstname.lastname@example.org
- Ph.D. Students
Beatriz Somovilla Crespo email@example.com
Isabel Corraliza Gorjón firstname.lastname@example.org
- Master Students
Lisa Kochsiek email@example.com
The CNB-CSIC Protein Tools Unit focuses on the design, production, characterization and use of proteins as specific molecular tools. The Unit generates monoclonal antibodies (mAb), performs studies of the immune response, develops specific immunoassays and analyzes biomolecular interactions using a Biacore 3000 surface plasmon resonance biosensor. It works in a horizontal manner, participating in research projects or on contract for specific goals. The activities, individually or as part of a project, are based on proposals from research groups within the CNB and from private or public research organizations and/or companies.
One of our main goals is to generate mAb not available commercially. These mAb, made for research purposes, can also be used for commercial applications. We also help basic research groups with technology transfer from research.
The Unit generated mAb against more than 60 different antigens, for use in basic and pre-clinical research, drug discovery, and diagnostics. We produced mAb to chromatin-associated proteins (Dido, ScoHET), centrosomal proteins (CAP350), immunoglobulins (canine IgE), blood proteins (coagulation Factor V, CD5L), neurodegenerative disease-related proteins (TAU, beta amyloid peptides), mitochondrial proteins (SCAM3), kinases (P38, PI3K, DGK, GSK3, PKD, SADB), viral antigens (TEGV, SARS-E, gp120) and other human pathogens (Candida famata, Anisakis spp).
Monoclonal antibody development and characterization
We have developed mAb for structure-function studies, antigen mapping, neutralization, quantitative assays, pharmacokinetics, and affinity determination of enzymes (alkaline phosphatase, RNAse), nucleases (CEL I), transcription factors (DREAM), hormones and their receptors (LH, FSH, TSH, hCG, hGH, hGH antagonist B2036, Pegvisomant, GHR), growth factors and their receptors (IGF-1, IGF-2, EGF, IGF-1R, EGFR), growth factor-binding proteins (IGFBP-1, IGFBP-3, GHBP), cytoskeletal proteins (4.1R, WIP), integral membrane proteins (MAL, BENE), blood proteins (albumin, immunoglobulins, Factor VIII), chemokines (CCL1, CCL11, CCL20), virus (Phi 29, torovirus) and tumor antigens (FBP). These mAb have been used by various international research groups and have been described in more than 200 scientific publications.
Production and characterization of antibodies to murine chemokine receptors using rats and knockout mice
We have generated a panel of poly- and monoclonal antibodies to chemokine receptors for studies of receptor-ligand interactions and for phenotyping mouse models of human inflammatory diseases. Some anti-mouse CCR4 and CCR8 mAb were produced by immunization of rats and knockout mice. Several of these mAb block ligand-receptor binding. One of them has been used by various research groups to inhibit CCR8 function in distinct mouse models of inflammatory disease and has been licensed to a company for worldwide commercialization.
Production and characterization of mAb to human chemokine receptors
We have generated a panel of mouse mAb to human chemokine receptors CCR6 and CCR9. One mAb against hCCR6 has been licensed for commercialization and two mAb specific for hCCR9 have been licensed as antitumor agents.
- Strategy design and implementation for immunization, immunogen production and analysis of animal immune responses
- Poly- and monoclonal antibody production in rabbit, mouse, rat and hamster
- Immunization with plasmids and transfected cells
- Antibody characterization by ELISA, RIA, flow cytometry, Western blot
- Purification and labeling of enzymes and antibodies
- Development of immunoassays for antigen detection and quantification in biological fluids
- Characterization of biomolecular interactions in real time using a Biacore 3000 biosensor robot. Determination of kinetic and affinity constants.
- Kremer L, Garcia-Sanz JA. Editorial: Is the Recent Burst of Therapeutic Anti-tumor Antibodies the Tip of an Iceberg? Front Immunol. 2018; 9: 442.
- Somovilla-Crespo B, Martín Monzón MT, Vela M, Corraliza-Gorjón I, Santamaria S, Garcia-Sanz JA, Kremer L. 92R Monoclonal Antibody Inhibits Human CCR9(+) Leukemia Cells Growth in NSG Mice Xenografts. Front Immunol. 2018; 9: 77.
- Cuesta-Mateos C, Alcaraz-Serna A, Somovilla-Crespo B, Muñoz-Calleja C. Monoclonal Antibody Therapies for Hematological Malignancies: Not Just Lineage-Specific Targets. Front Immunol. 2018; 8: 1936.
- Corraliza-Gorjón I, Somovilla-Crespo B, Santamaria S, Garcia-Sanz JA, Kremer L. New Strategies Using Antibody Combinations to Increase Cancer Treatment Effectiveness. Front Immunol. 2017; 8: 1804.
- Pose-Utrilla J, García-Guerra L, Del Puerto A, Martín A, Jurado-Arjona J, De León-Reyes NS, Gamir-Morralla A, Sebastián-Serrano Á, García-Gallo M, Kremer L, Fielitz J, Ireson C, Pérez-Álvarez MJ, Ferrer I, Hernández F, Ávila J, Lasa M, Campanero MR, Iglesias T. Excitotoxic inactivation of constitutive oxidative stress detoxification pathway in neurons can be rescued by PKD1. Nat Commun. 2017; 8: 2275.
- Santamaria S, Delgado M, Kremer L, Garcia-Sanz JA. Will a mAb-Based Immunotherapy Directed against Cancer Stem Cells Be Feasible? Front Immunol. 2017; 8: 1509.
- Roncador G, Engel P, Maestre L, Anderson AP, Cordell JL, Cragg MS, Šerbec VC, Jones M, Lisnic VJ, Kremer L, Li D, Koch-Nolte F, Pascual N, Rodríguez-Barbosa JI, Torensma R, Turley H, Pulford K, Banham AH. The European antibody network's practical guide to finding and validating suitable antibodies for research. MAbs. 2016; 8: 27-36.
- Bartolini F, Andres-Delgado L, Qu X, Nik S, Ramalingam N, Kremer L, Alonso MA, Gundersen GG. An mDia1-INF2 formin activation cascade facilitated by IQGAP1 regulates stable microtubules in migrating cells. Mol Biol Cell. 2016; 27: 1797-808.
- Ruiz MF, Alvarez M, Eirín-López JM, Sarno F, Kremer L, Barbero JL, Sánchez L. An Unusual Role for doublesex in Sex Determination in the Dipteran Sciara. Genetics. 2015, 200: 1181-99.
- Vela M, Aris M, Llorente M, Garcia-Sanz JA, Kremer L. Chemokine receptor-specific antibodies in cancer immunotherapy: achievements and challenges. Front Immunol 2015; 6: 2.
- González-Magaldi M, Martín-Acebes MA, Kremer L, Sobrino F. Membrane topology and cellular dynamics of foot-and-mouth disease virus 3A protein. PLoS One. 2014, 9: e106685.
- Barbarroja-Escudero J, Prieto-Martin A, Monserrat-Sanz J, Reyes-Martin E, Diaz-Martin D, Antolin-Amerigo D, Rodriguez-Rodriguez M, Canseco-Gonzalez F, Kremer L, Martinez-A C, Alvarez-Mon M. Abnormal chemokine receptor profile on circulating T lymphocytes from nonallergic asthma patients. Int Arch Allergy Immunol. 2014, 164: 228-36.
- Goñi GM, Epifano C, Boskovic J, Camacho-Artacho M, Zhou J, Bronowska A, Martín MT, Eck MJ, Kremer L, Gräter F, Gervasio FL, Perez-Moreno M, Lietha D. Phosphatidylinositol 4,5-bisphosphate triggers activation of focal adhesion kinase by inducing clustering and conformational changes. Proc Natl Acad Sci U S A. 2014, 111: E3177-86.
- Chamorro S, Vela M, Franco-Villanueva A, Carramolino L, Gutiérrez J, Gómez L, Lozano M, Salvador B, García-Gallo M, Martínez-A C, Kremer L. Antitumor effects of a monoclonal antibody to human CCR9 in leukemia cell xenografts. MAbs. 2014; 6: 1000-12.
- Aranda JF, Reglero-Real N, Marcos-Ramiro B, Ruiz-Sáenz A, Fernández-Martín L, Bernabé-Rubio M, Kremer L, Ridley AJ, Correas I, Alonso MA, Millán J. MYADM controls endothelial barrier function through ERM-dependent regulation of ICAM-1 expression. Mol Biol Cell. 2013; 24: 483-94.
- Sánchez-Martín D, Martínez-Torrecuadrada J, Teesalu T, Sugahara KN, Alvarez-Cienfuegos A, Ximénez-Embún P, Fernández-Periáñez R, Teresa Martín M, Molina-Privado I, Ruppen-Cañás I, Blanco-Toribio A, Cañamero M, Cuesta AM, Compte M, Kremer L, Bellas C, Alonso-Camino V, Guijarro-Muñoz I, Sanz L, Ruoslahti E, Alvarez-Vallina L. Proteasome activator complex PA28 identified as an accessible target in prostate cancer by in vivo selection of human antibodies. Proc Natl Acad Sci U S A. 2013, 110: 13791-6.
- Somovilla-Crespo B, Alfonso-Pérez M, Cuesta-Mateos C, Carballo-de Dios C, Beltrán AE, Terrón F, Pérez-Villar JJ, Gamallo-Amat C, Pérez-Chacón G, Fernández-Ruiz E, Zapata JM, Muñoz-Calleja C. Anti-CCR7 therapy exerts a potent anti-tumor activity in a xenograft model of human mantle cell lymphoma. J Hematol Oncol. 2013; 6: 89.
- Inventors: Chamorro S, Franco A, García Sanz JA, Kremer L, Martínez Alonso C, Vela Cuenca M, Carramolino L. “Antibodies against CCR9 and applications thereof”. EP13382469.8, PCT/EP2014/075578 (2014), CSIC.
- Inventors: Kremer L, Carrasco Llamas L, Pisa García D. “Monoclonal antibody against the protein GAPDH from Candida famata”. 200930966 (2009), CSIC and Universidad Autónoma Foundation.
- Inventors: Kremer L, Llorente M, Casasnovas JM, Fernández Ruiz E, Galán Diez M. “Antibody anti-dectin-1, hybridoma producer of this antibody and its applications”. P200801766 (2008), CSIC and Biomedical Research Foundation of the La Princesa Hospital.
- Inventors: Bernard A, Garmendia C, Abad JL, Gonzalez MA, Llorente M, Martinez-A C, Serrano F. “Multifunctional genetic constructions having a high capacity to inhibit the expression of CCR5 on the cell surface”. WO2004013330 (2004), Genetrix SL and CSIC.
- Inventors: Kremer L, Mellado JM, Rodríguez Frade JM, Martínez Alonso C. “Method for determining cytokine receptor activation by the use of an antibody”. WO9841872, CA2285036, AU6530498, US2003077663, PCT/SE98/00496 (1998 & 2003), Pharmacia SA and CSIC.
- Inventors: Llorente M, Mendez E, Lopez E. “Method for extracting gluten contained in heat-processed and non-heat-processed foodstuffs, compatible with an enzyme-linked immunosorbent assay, composition and kits comprising said composition”. WO02092633 (2002), CSIC.
- Inventors: Kremer L, Mellado JM, Rodríguez-Frade JM, Martínez Alonso C, Hansson YE. “Monoclonal antibodies binding human growth hormone (hGH)”. SE 9601231-5, W09736929, NZ331868, AU2525597, US5945296 (1997), Pharmacia AB.
- CroatiaStipan JonjicUniversity of Rijeka
- DenmarkJakub Z. KaczmarekSanovo Biotech A/S
- FranceAndré PèlegrinInstitut de Recherche en Cancérologie de Montpellier, IRCM
- FranceArmand BensussanHôpital Saint Louis
- FrancePierre LafayeInstitut Pasteur
- GermanyFriedrich Koch-NolteUniversity Medical Center Hamburg-Eppendorf
- HungaryJohanna SymmonsEnglish & Scientific Consulting Kft. (SCICONS)
- HungaryPeter BaloghClinical Center, University of Pecs
- ItalyFabio MalavasiUniversity of Torino Medical School
- RussiaAlexander FilatovInstitute of Immunology
- RussiaAlexander TaraninInstitute of Molecular and Cellular Biology
- SloveniaVladka Čurin ŠerbecBlood Trasfusion Centre of Slovenia
- SpainGiovanna RoncadorCentro Nacional de Investigaciones Oncológicas
- SpainJosé I. Rodríguez BarbosaEdificio Nuevos Institutos de Investigación
- SpainLeonor KremerCentro Nacional de Biotecnología/CSIC
- SpainNúria PascualInstitute for Advanced Chemistry of Catalonia (IQAC-CSIC)
- SpainPablo EngelFaculty of Medicine and Medical Sciences, University of Barcelona
- United KingdomUniversity of OxfordLRF Haemato-oncology Group
- United KingdomVertebrate Antibodies Limited (VAL)
- United KingdomKing's College LondonSt. John's Institute of Dermatology