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

Juan Carlos Barragan-Galvez, Maria Gonzalez-Orozco, Araceli Hernandez-Flores, Jose Luis Maravillas-Montero, Yedhani Chavez-Guerrero, Vianney Ortiz-Navarrete

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)224-231
Number of pages8
JournalProtein Journal
Volume39
Issue number3
DOIs
StatePublished - Jun 1 2020

Keywords

  • CRTAM
  • Nectin-like 2
  • Recombinant protein
  • Superfamily of immunoglobulins

ASJC Scopus subject areas

  • Analytical Chemistry
  • Bioengineering
  • Biochemistry
  • Organic Chemistry

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  • Cite this

    Barragan-Galvez, J. C., Gonzalez-Orozco, M., Hernandez-Flores, A., Maravillas-Montero, J. L., Chavez-Guerrero, Y., & Ortiz-Navarrete, V. (2020). Prokaryotic Expression of the Immunoglobulin’s Domains of CRTAM to Characterize a Monoclonal Antibody. Protein Journal, 39(3), 224-231. https://doi.org/10.1007/s10930-020-09896-y