Abstract
C-reactive protein (CRP) is a pentameric protein that is present in the bloodstream during inflammatory events, e.g., liver failure, leukemia, and/or bacterial infection. The level of CRP indicates the progress and prognosis of certain diseases; it is therefore necessary to measure CRP levels in the blood accurately. The normal concentration of CRP is reported to be 1–3 mg/L. Inflammatory events increase the level of CRP by up to 500 times; accordingly, CRP is a biomarker of acute inflammatory disease. In this study, we demonstrated the preparation of DNA aptamer-conjugated peripheral blood mononuclear cells (Apt-PBMCs) that specifically capture human CRP. Live PBMCs functionalized with aptamers could detect different levels of human CRP by producing immune complexes with reporter antibody. The binding behavior of Apt-PBMCs toward highly concentrated CRP sites was also investigated. The immune responses of Apt-PBMCs were evaluated by measuring TNF-alpha secretion after stimulating the PBMCs with lipopolysaccharides. In summary, engineered Apt-PBMCs have potential applications as live cell based biosensors and for in vitro tracing of CRP secretion sites.
Highlights
C-reactive protein (CRP) is a pentameric protein that is present in the bloodstream during inflammatory events, e.g., liver failure, leukemia, and/or bacterial infection
Controlling the optimal amount of calcium ions in the cell culture and reagents is a key factor in CRP detection using Apt-Peripheral blood mononuclear cells (PBMCs)
To prove our hypothesis that each PBMC may function as a cell sensor, we introduced magnetic beads as cell cores
Summary
C-reactive protein (CRP) is a pentameric protein that is present in the bloodstream during inflammatory events, e.g., liver failure, leukemia, and/or bacterial infection. We demonstrated the preparation of DNA aptamer-conjugated peripheral blood mononuclear cells (Apt-PBMCs) that capture human CRP. Live PBMCs functionalized with aptamers could detect different levels of human CRP by producing immune complexes with reporter antibody. Engineered Apt-PBMCs have potential applications as live cell based biosensors and for in vitro tracing of CRP secretion sites. Immunochemical techniques, depend on the quality of antibodies for their sensitivity and specificity, which limits their applications[1,2]. C-reactive protein (CRP) is a ring-shaped, pentameric protein produced in the liver; it increases in the serum upon infection or immunological response, and is especially upregulated in the case of cardiovascular disease[11,12]. CRP, we prepared the CRP standards in their concentration ranges from 0.01 to 30 mg/L, which covers the usual ranges of CRP concentrations specific for heart disease or inflammations (1–10 mg/L)[11]
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