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

Barragan-Galvez, Juan Carlos; González-Orozco, María; Hernandez-Flores, Araceli; Maravillas-Montero, José Luis; Chavez-Guerrero, Yedhani; Ortı́z-Navarrete, Vianney

doi:10.1007/s10930-020-09896-y

Cited by 2 publications

(4 citation statements)

References 21 publications

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“…We previously produced the variable domain (CRTAM IgV−stalk ), the constant domain (CRTAM IgC−stalk ), and the extracellular region of CRTAM (CRTAM EC ) as recombinant proteins [15]. However, we noticed these recombinant proteins formed in aggregates with molecular weights greater than 2,000 kDa (Supplementary Fig.…”

Section: Resultsmentioning

confidence: 99%

“…1: CRTAM EC , CRTAM IgV−Stalk , CRTAM IgC−Stalk and NECL2 EC . These constructions were cloned into a pET28a (+) expression vector (Novagen) as described previously [15]. Other constructions we designed for mammalian expression: CRTAM full , CRTAM IgV , CRTAM IgC and NECL2 full .…”

Section: Methodsmentioning

confidence: 99%

“…Expression and puri cation of recombinant proteins were done as previously described [15]. The puri ed proteins were subjected to a chemical refolding as described by Zhang [17].…”

Section: Protein Expression and Puri Cationmentioning

confidence: 99%

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The constant domain of CRTAM is essential for high-affinity interaction with Nectin-like 2

Barragan-Galvez,

Lopez-Ortega,

Hernandez-Flores

et al. 2023

Preprint

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CRTAM (Class-I MHC-restricted T cell-associated molecule) is a member of the Nectin-like family, composed of two extracellular domains, one constant domain (IgC), and another variable domain (IgV), expressed in activated CD8 T cells, epithelial cells, NK cells and in a subpopulation CD4 T cell. CRTAM recognizes the ligand Nectin-like 2 (Necl2) through the variable domain. However, the role of the constant domain during this ligand recognition has yet to be understood. In this study, we show the purification of soluble-folded Ig-domains of CRTAM, and we demonstrated that the constant domain forms a homodimer in solution via hydrophobic interactions. By SRP analysis, we also demonstrate that the full CRTAM binds to Necl2 with a 2.16 nM affinity. Additionally, transfected cells expressing the full length of CRTAM on the surface establish a better interaction with Necl2 than with the CRTAM-IgC domain expressed on the cell surface. In conclusion, the constant domain of CRTAM is essential for high-affinity interaction with Necl-2.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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The constant domain of CRTAM is essential for high-affinity interaction with Nectin-like 2

Barragan-Galvez,

Lopez-Ortega,

Hernandez-Flores

et al. 2023

Preprint

Self Cite

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“…The occurrence of severe pneumonia is associated with factors such as susceptibility, immune insufficiency, and weak resistance [9][10][11][12]. Immunoglobulins are immune agents extracted from plasma that, when administered intravenously, neutralize pathogenic bacteria and modulate the immune and inflammatory response in patients with infectious diseases [13,14].…”

Section: Introductionmentioning

confidence: 99%

Human immunoglobulin in combination with antimicrobial agents enhances the treatment efficacy and reduces inflammatory response in children with severe pneumonia

Zhang

2024

Am J Transl Res

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Objective: To investigate the efficacy of human immunoglobulin combined with antibiotics in treating severe pediatric pneumonia. Methods: A retrospective analysis was performed on 210 pediatric patients with severe pneumonia admitted to the Department of Neonatology of Cangzhou Central Hospital from April 2019 to October 2022. Patients were divided into two groups (the observation group and the control group) based on the administration of human immunoglobulin. Clinical indexes of both groups before and after treatment were analyzed to determine the therapeutic effect of different treatment methods on pediatric severe pneumonia. Results: The durations of cough, fever, pulmonary rales, and lung shadow, and hospitalization time in the observation group were significantly shorter than those in the control group (all P<0.05). The total clinical effective rate in the observation group was significantly higher than that in the control group (P<0.05). Levels of inflammatory factors (IL-6, IL-8 and hsCRP) were decreased in both groups after treatment (all P<0.05), and were lower in the observation group compared with the control group after treatment (all P<0.05). The serum levels of IgA, IgG and IgM after five days of intervention were obviously higher than those before intervention in the observation group (all P<0.05), but the serum levels of IL-4, INF-γ and INF-γ/IL-4 were obviously lower (all P<0.05). The total incidence of adverse reactions between two groups after intervention was not statistically different (P<0.05). Conclusion: The combination of human immunoglobulin and antibiotics for the treatment of pediatric severe pneumonia is beneficial, because it improves efficacy, boosts the immune system, and reduces inflammation.

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

Cited by 2 publications

References 21 publications

The constant domain of CRTAM is essential for high-affinity interaction with Nectin-like 2

The constant domain of CRTAM is essential for high-affinity interaction with Nectin-like 2

Human immunoglobulin in combination with antimicrobial agents enhances the treatment efficacy and reduces inflammatory response in children with severe pneumonia

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