TY - JOUR
T1 - Prokaryotic Expression of the Immunoglobulin’s Domains of CRTAM to Characterize a Monoclonal Antibody
AU - Barragan-Galvez, Juan Carlos
AU - Gonzalez-Orozco, Maria
AU - Hernandez-Flores, Araceli
AU - Maravillas-Montero, Jose Luis
AU - Chavez-Guerrero, Yedhani
AU - Ortiz-Navarrete, Vianney
N1 - Publisher Copyright:
© 2020, Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2020/6/1
Y1 - 2020/6/1
N2 - 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.
AB - 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.
KW - CRTAM
KW - Nectin-like 2
KW - Recombinant protein
KW - Superfamily of immunoglobulins
UR - http://www.scopus.com/inward/record.url?scp=85083504176&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85083504176&partnerID=8YFLogxK
U2 - 10.1007/s10930-020-09896-y
DO - 10.1007/s10930-020-09896-y
M3 - Article
C2 - 32300914
AN - SCOPUS:85083504176
SN - 1572-3887
VL - 39
SP - 224
EP - 231
JO - Protein Journal
JF - Protein Journal
IS - 3
ER -