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Prokaryotic Expression of the Immunoglobulin’s Domains of CRTAM to Characterize a Monoclonal Antibody

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Abstract

Class-I restricted T cell-associated molecule (CRTAM) is a member of the immunoglobulin superfamily, and it is closely related to nectin-like protein. CRTAM is expressed in activated CD8 T cells, NKT cells, NK cells and in a subpopulation CD4 T cells. In this study, we produce as recombinant proteins, the Ig-domains of CRTAM (IgV–IgC), the IgV, and the IgC. These proteins were successfully purified in the soluble fraction only if the stalk region was included. The recombinant CRTAM recognizes its ligand nectin-like 2 in a cell-free system. We also demonstrate that the IgC domain of CRTAM is recognized by the anti-hCRTAM monoclonal antibody C8 with a 0.62 nM affinity. In conclusion, the stalk region of CRTAM provides solubility for the expression of its Ig-domains as recombinant proteins.

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Funding

The present work was supported by Cinvestav Grant to Vianney Ortiz Navarrete. Barragan-Galvez Juan Carlos is a doctoral student from the Programa de Doctorado en Ciencias Biomedicas, Universidad Nacional Autónoma de México (UNAM) and was supported by Consejo Nacional de Ciencia y Tecnología (CONACYT) scholarship 489981. The funding agencies had no role in study design, data collection and analysis, preparation of the manuscript or the decision to publish.

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Authors and Affiliations

  1. Department of Molecular Biomedicine, Center for Research and Advanced Studies (CINVESTAV), Av. IPN 2508. Col. San Pedro Zacatenco, Gustavo A Madero, 07360, Mexico City, Mexico

    Maria Gonzalez-Orozco, Yedhani Chavez-Guerrero & Vianney Ortiz-Navarrete

  2. Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, USA

    Maria Gonzalez-Orozco

  3. Universidad de la Sierra Sur, Miahuatlán de Porfirio Díaz, Oaxaca, Mexico

    Araceli Hernandez-Flores

  4. Research Support Network, Universidad Nacional Autónoma de México and Instituto Nacional de Ciencias Médicas y Nutrición “Salvador Zubirán”, Mexico City, Mexico

    Jose Luis Maravillas-Montero

  5. Biomedical Sciences Program, National Autonomous University of Mexico, Circuito Interior s/n, Cd. Universitaria, Mexico City, Mexico

    Juan Carlos Barragan-Galvez

Authors
  1. Juan Carlos Barragan-Galvez
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  2. Maria Gonzalez-Orozco
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  3. Araceli Hernandez-Flores
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  4. Jose Luis Maravillas-Montero
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  5. Yedhani Chavez-Guerrero
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  6. Vianney Ortiz-Navarrete
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Correspondence to Vianney Ortiz-Navarrete.

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Barragan-Galvez, J.C., Gonzalez-Orozco, M., Hernandez-Flores, A. et al. Prokaryotic Expression of the Immunoglobulin’s Domains of CRTAM to Characterize a Monoclonal Antibody. Protein J 39, 224–231 (2020). https://doi.org/10.1007/s10930-020-09896-y

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  • DOI: https://doi.org/10.1007/s10930-020-09896-y

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