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
- Lab Members
Mónica García-Gallo (PhD) email@example.com
María Teresa Martín (PhD) firstname.lastname@example.org
Mercedes Llorente (BSc) email@example.com
Laura Martín (BSc) firstname.lastname@example.org
Carolina Sánchez email@example.com
Lucio Gómez firstname.lastname@example.org
The CNB-CSIC Protein Tools Unit focuses on the design, creation 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.
Our main goal is to generate mAb not available commercially. These mAb, made for research purposes, can also be used for patent and commercial applications; we also help basic research groups with technology transfer from research. In its first two years of activity (2007-2008), the Unit generated nearly 100 mAb against more than 30 antigens, for use in basic and pre-clinical research, drug discovery, and diagnostics. We currently produce mAb to chromatin-associated proteins (Dido, ScoHET), kinases (P38, PI3K, DGK, GSK3, PKD), receptors (Dectin-1, Kidins220), viral antigens (TEGV, SARS) 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 (LH, FSH, TSH, hCG, hGH, hGH antagonist B2036, Pegvisomant), growth factors (IGF-1, IGF-2, EGF), growth factor-binding proteins (IGFBP-1, IGFBP-3, GHBP), cytoskeletal proteins (4.1R, WIP), receptors (IGF-1R, EGFR, GHR, CCR4, CCR6, CCR8, CCR9), integral membrane proteins (MAL, BENE), plasma and coagulation 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 of which recognizes hCCR6 in flow cytometry and has been licensed for commercialization.
Production of poly- and monoclonal antibodies to cell signaling molecules
Our current research focuses on phosphospecific antibodies against kinases, including glycogen synthase kinase 3 (GSK3), protein kinase D (PKD), class I(A) phosphoinositide 3-kinase (PI3K), SADB Ser/Thr kinase, diacylglycerol kinase (DGK) and a specific epitope in the p38 MAP kinase (MAPK) alternative pathway.
- 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.
- 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.
- González-Magaldi M, Postigo R, de la Torre BG, Vieira YA, Rodríguez-Pulido M, López-Viñas E, Gómez-Puertas P, Andreu D, Kremer L, Rosas MF, Sobrino F. Mutations that hamper dimerization of foot-and-mouth disease virus 3A protein are detrimental for infectivity. J Virol. 2012; 86: 11013-23.
- Cortés I, Sánchez-Ruíz J, Zuluaga S, Calvanese V, Marqués M, Hernández C, Rivera T, Kremer L, González-García A, Carrera AC. p85β phosphoinositide 3-kinase subunit regulates tumor progression. Proc Natl Acad Sci U S A. 2012; 109: 11318-23.
- Inventors: Kremer L, Llorente M, Casasnovas JM, Fernández Ruiz E, Galán Diez M. “Anticuerpo anti-Dectin-1, hibridoma productor de dicho anticuerpo y sus aplicaciones”. P200801766, 2008. CSIC y Fundación para la Investigación Biomédica del Hospital de La Princesa.
- 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
- 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 PascualNb4D
- SpainPablo EngelMedical School, University of Barcelona
- United KingdomUniversity of OxfordLRF Haemato-oncology Group
- United KingdomVertebrate Antibodies Limited (VAL)
- United KingdomUniversity of OxfordOxFabs
- United KingdomKing's College LondonSt. John's Institute of Dermatology