Browsing by Subject "enzyme linked immunosorbent assay"

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    Assessment of diagnostic enzyme-linked immunosorbent assay kit and serological markers in human brucellosis
    (2008) Cakan G.; Bezirci F.B.; Kacka, A.; Cesur, S.; Aksaray, S.; Tezeren, D.; Saka, D.; Ahmed, K.
    This study was performed to evaluate commercial brucella immunoglobulin G and M-enzymelinked immunosorbent assay (IgG and IgM ELISA) kits for the diagnosis of human brucellosis and to suggest a candidate prognostic marker for human brucellosis. We determined the serum levels of brucella IgG, IgM, C-reactive protein (CRP), soluble CD14 (sCD 14), and neopterin in patients with brucellosis and compared them with those of normal healthy persons, patients with tuberculosis, and patients with other diseases. It was found that the sensitivity of ELISA to diagnose brucellosis was high when both IgG and IgM ELISA were used together. This study showed that serum CRP, sCD14, or neopterin levels were significantly high during the course of human brucellosis. The above markers, alone or in combination, might have the potential to evaluate treatment outcomes in human brucellosis. The markers that can predict the variability of agglutination titer was also determined. It was found that the titer value alone does not fully represent disease status.
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    Identification of novel neutralizing single-chain antibodies against vascular endothelial growth factor receptor 2
    (2011) Erdag, B.; Koray Balcioglu, B.; Ozdemir Bahadir, A.; Serhatli, M.; Kacar O.; Bahar, A.; Seker, U.O.S.; Akgun, E.; Ozkan, A.; Kilic, T.; Tamerler, C.; Baysal, K.
    Human vascular endothelial growth factor (VEGF) and its receptor (VEGFR-2/kinase domain receptor [KDR]) play a crucial role in angiogenesis, which makes the VEGFR-2 signaling pathway a major target for therapeutic applications. In this study, a single-chain antibody phage display library was constructed from spleen cells of mice immunized with recombinant human soluble extracellular VEGFR-2/KDR consisting of all seven extracellular domains (sKDR D1-7) to obtain antibodies that block VEGF binding to VEGFR-2. Two specific single-chain antibodies (KDR1.3 and KDR2.6) that recognized human VEGFR-2 were selected; diversity analysis of the clones was performed by BstNI fingerprinting and nucleotide sequencing. The single-chain variable fragments (scFvs) were expressed in soluble form and specificity of interactions between affinity purified scFvs and VEGFR-2 was confirmed by ELISA. Binding of the recombinant antibodies for VEGFR-2 receptors was investigated by surface plasmon resonance spectroscopy. In vitro cell culture assays showed that KDR1.3 and KDR2.6 scFvs significantly suppressed the mitogenic response of human umbilical vein endothelial cells to recombinant human VEGF 165 in a dose-dependent manner, and reduced VEGF-dependent cell proliferation by 60% and 40%, respectively. In vivo analysis of these recombinant antibodies in a rat cornea angiogenesis model revealed that both antibodies suppressed the development of new corneal vessels (p < 0.05). Overall, in vitro and in vivo results disclose strong interactions of KDR1.3 and KDR2.6 scFvs with VEGFR-2. These findings indicate that KDR1.3 and KDR2.6 scFvs are promising antiangiogenic therapeutic agents. © 2011 International Union of Biochemistry and Molecular Biology, Inc.
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    Inflammasome induction and immunostimulatory effects of CpG-ODN loaded liposomes containing DC-cholesterol
    (Turkish Society of Immunology, 2013) Bayyurt, B.; Gürsel I.
    Objectives: This study aims to investigate the effects of cholesterol content and cationic property of liposomes on immune response. Materials and methods: Liposomes containing high amounts of 3β-[N-(N',N'-dimethylaminoethane)-carbamoyl] cholesterol hydrochloride (DC-cholesterol) were prepared and loaded with K- and D-type CpG oligonucleotide (CpG-ODN) via dehydration-rehydration (DRV) method. After splenocytes and peritoneal exudate cells (PECs) primed with lipopolysaccharide (LPS) was incubated either with free or liposomal CpG-ODN counterparts, supernatants were collected and used in cytokine (IFN-g, IL-1γ and IL-1β) ELISA. Additionally, supernatants of PECs primed with LPS and stimulated with liposomes containing different doses of DC-cholesterol were collected and used in IL-1β ELISA assay. Results: Low-dose CpG-ODN loaded liposomal formulations induced higher immune activation than free CpG-ODN at the same dose. While high-dose liposomal CpG-ODN formulations decreased pro-inflammatory cytokine production in splenocytes, they increased the secretion of IL-1β. Inflammasome activation was increased in a dose dependent manner when PECs primed with LPS were incubated with only liposomes. Varying lipid molar ratios of DC-Cholesterol containing liposomes increased IL-1β production based on increasing lipid molar ratio. Conclusion: This study revealed that type and lipid ratio of liposomes may alter the cellular efficacy of the loaded immune-stimulatory agent and may initiate inflammasome activation. © 2014 Turkish Journal of Immunology.
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    MAb 6D5 against proteins overexpressed in hepatocellular carcinoma cell lines
    (2007) Yagci, T.
    [No abstract available]