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Prokaryotic Expression of the Immunoglobulin's Domains of CRTAM to Characterize a Monoclonal Antibody.
The Protein Journal ( IF 3 ) Pub Date : 2020-04-16 , DOI: 10.1007/s10930-020-09896-y
Juan Carlos Barragan-Galvez 1 , Maria Gonzalez-Orozco 2, 3 , Araceli Hernandez-Flores 4 , Jose Luis Maravillas-Montero 5 , Yedhani Chavez-Guerrero 2 , Vianney Ortiz-Navarrete 2
Affiliation  

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.

中文翻译:

CRTAM免疫球蛋白结构域的原核表达,以表征单克隆抗体。

I类限制性T细胞相关分子(CRTAM)是免疫球蛋白超家族的成员,并且与类似果胶的蛋白密切相关。CRTAM在活化的CD8 T细胞,NKT细胞,NK细胞和亚群CD4 T细胞中表达。在这项研究中,我们生产CRTAM(IgV–IgC),IgV和IgC的Ig结构域作为重组蛋白。仅在包括茎区域的情况下,这些蛋白才能在可溶性部分中成功纯化。重组CRTAM在无细胞系统中识别其配体类似nectin的2。我们还证明了CRTAM的IgC域被抗hCRTAM单克隆抗体C8识别为0.62 nM亲和力。总之,CRTAM的茎区域为重组蛋白表达其Ig结构域提供了溶解性。
更新日期:2020-04-16
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