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Leonor Kremer

Name of the laboratory

CNB-CSIC Protein Tools Unit
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) lkremer@cnb.csic.es
Technicians
Mónica García-Gallo (PhD) mggallo@cnb.csic.es
María Teresa Martín (PhD) mtmartin@cnb.csic.es
Ana María García Cabrero (PhD) amgarcia@cnb.csic.es
Mercedes Llorente (BSc) llorente@cnb.csic.es
Ph.D. Students
Beatriz Somovilla Crespo bsomovilla@cnb.csic.es
Isabel Corraliza Gorjón icorraliza@cnb.csic.es
Master Students
Lisa Kochsiek lisa.kochsiek@uni-bielefeld.de

Laboratory activity

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).

Research activities

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.

Techniques available

  • 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.

Publications (2018-present)

  • 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.

Patents

  • 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.