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Antibody Array (antibody + array)
Selected AbstractsAttovial-based antibody nanoarraysPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 24 2009Peter Ellmark Abstract Antibody array-based technology is a powerful emerging tool in proteomics, but to enable global proteome analysis, antibody array layouts with even higher density has to be developed. To this end, we have further developed the first generation of a nanoarray platform, based on attoliter-sized vials, attovials, which we have characterized and used for the detection of complement factor C1q in human serum samples. Finally, we demonstrated proof-of-concept for individual functionalization of the attovials with a recombinant antibody. [source] Simple Fabrication of Antibody Microarrays on Nonfouling Polymer Brushes with Femtomolar Sensitivity for Protein Analytes in Serum and BloodADVANCED MATERIALS, Issue 19 2009Angus Hucknall A multianalyte antibody array that is spotted on a poly(oligo(ethylene glycol) methacrylate) brush 100,nm thick, grown on glass via surface-initiated atom transfer radical polymerization, has femtomolar limit-of-detection (LOD) of cytokines in serum and whole blood, and a dynamic range of six orders of magnitude for a range of protein analytes. [source] Hyperosmotic stress induces Axl activation and cleavage in cerebral endothelial cellsJOURNAL OF NEUROCHEMISTRY, Issue 1 2008Imola Wilhelm Abstract Because of the relative impermeability of the blood-brain barrier (BBB), many drugs are unable to reach the CNS in therapeutically relevant concentration. One method to deliver drugs to the CNS is the osmotic opening of the BBB using mannitol. Hyperosmotic mannitol induces a strong phosphorylation on tyrosine residues in a broad spectrum of proteins in cerebral endothelial cells, the principal components of the BBB. Previously, we have shown that among targets of tyrosine phosphorylation are ,-catenin, extracellular signal-regulated kinase 1/2 and the non-receptor tyrosine kinase Src. The aim of this study was to identify new signalling pathways activated by hypertonicity in cerebral endothelial cells. Using an antibody array and immunoprecipitation we identified the receptor tyrosine kinase Axl to become tyrosine phosphorylated in response to hyperosmotic mannitol. Besides activation, Axl was also cleaved in response to osmotic stress. Degradation of Axl proved to be metalloproteinase- and proteasome-dependent and resulted in 50,55 kDa C-terminal products which remained phosphorylated even after degradation. Specific knockdown of Axl increased the rate of apoptosis in hyperosmotic mannitol-treated cells; therefore, we assume that activation of Axl may be a protective mechanism against hypertonicity-induced apoptosis. Our results identify Axl as an important element of osmotic stress-induced signalling. [source] Antibody microarray analysis of serum glycans in esophageal squamous cell carcinoma cases and controlsPROTEOMICS - CLINICAL APPLICATIONS, Issue 8 2009Changxia Shao Abstract The objective of this study was to characterize specific serum glycan profile in esophageal squamous cell carcinoma (ESCC) cases and matched controls. A recently developed lectin-based antibody array was applied to detect various cancer-associated glycotopes in sera from 23 cases and 23 controls. Glycan levels were highly expressed in sera of ESCC patients as compared with controls. These included fucosylation level on interleukin (IL) 8; mannose level on haptoglobin; N-acetylglucosamine levels on IL6, mucin (MUC)1, and von Willebrand factor (vWf); sialyl Lewis a (sLea) levels on blood group Lewis X, IL6, IL10, MUC1, and serum amyloid A (SAA); sialyl Tn antigen (sTn) levels on cathepsin D, gelsolin, IL10, and vWf; T antigen levels on IL8, IL10, blood group Lewis X, vitronectin, and vWf (p<0.05). In addition, receiver-operating characteristics (ROC) analysis showed significantly discriminal improvement between cases and controls as measured by area under ROC curve. The highest sum of sensitivity and specificity was 1.52 for carbohydrate antigen 19-9 detection on both MUC1 and SAA, with area under ROC curves of 0.73 and 0.71, respectively. Taken together, this lectin-based antibody microarray allows efficient profiling of variations in specific glycans on proteins in ESCC cases as compared with controls, some of which might be useful for clinical diagnosis, early detection, and/or treatment. [source] Blockade of IL-15 activity inhibits microglial activation through the NF,B, p38, and ERK1/2 pathways, reducing cytokine and chemokine releaseGLIA, Issue 3 2010Diego Gomez-Nicola Abstract Reactive glia formation is one of the hallmarks of damage to the CNS, but little information exists on the signals that direct its activation. Microglial cells are the main regulators of both innate and adaptative immune responses in the CNS. The proinflammatory cytokine IL-15 is involved in regulating the response of T and B cells, playing a key role in regulating nervous system inflammatory events. We have used a microglial culture model of inflammation induced by LPS and IFN, to evaluate the role of IL-15 in the proinflammatory response. Our results indicate that IL-15 is necessary for the reactive response, its deficiency (IL-15-/-) leading to the development of a defective proinflammatory response. Blockade of IL-15, both with blocking antibodies or with the ganglioside Neurostatin, inhibited the activation of the NF,B pathway, decreasing iNOS expression and NO production. Inhibiting IL-15 signaling also blocked the activation of the mitogen-activated protein kinase (MAPK) pathways ERK1/2 and p38. The major consequence of these inhibitory effects, analyzed using cytokine antibody arrays, was a severe decrease in the production of chemokines, cytokines and growth factors, like CCL17, CCL19, IL-12, or TIMP-1, that are essential for the development of the phenotypic changes of glial activation. In conclusion, activation of the IL-15 system seems a necessarystep for the development of glial reactivity and the regulation of the physiology of glial cells. Modulating IL-15 activity opens the possibility of developing new strategies to control gliotic events upon inflammatory stimulation. © 2009 Wiley-Liss, Inc. [source] Biomedical applications of protein chipsJOURNAL OF CELLULAR AND MOLECULAR MEDICINE, Issue 3 2002Jocelyn H. Ng Abstract The development of microchips involving proteins has accelerated within the past few years. Although DNA chip technologies formed the precedent, many different strategies and technologies have been used because proteins are inherently a more complex type of molecule. This review covers the various biomedical applications of protein chips in diagnostics, drug screening and testing, disease monitoring, drug discovery (proteomics), and medical research. The proteomics and drug discovery section is further subdivided to cover drug discovery tools (on-chip separations, expression profiling, and antibody arrays), molecular interactions and signaling pathways, the identification of protein function, and the identification of novel therapeutic compounds. Although largely focused on protein chips, this review includes chips involving cells and tissues as a logical extension of the type of data that can be generated from these microchips. [source] Protein screening in vitreous samples of patients with retinal vein occlusionACTA OPHTHALMOLOGICA, Issue 2009HT AGOSTINI Purpose The aim of the study was to identify proteins involved in the pathogenesis of retinopathy after retinal vein occlusion. In retinal vein occlusion, proteins penetrate from leaky vessels into the vitreous. Alternatively, retinal cells produce protein factors and release them into the vitreous. Methods Vitreous and plasma samples of patients with retinal vein occlusion or macular pucker / macular hole were analyzed by antibody microarrays and ELISA. Results An antibody based microarray with more than 500 target for screening vitreous samples initially was less enlightening than antibody arrays providing the possibility to quantify up to 30 proteins in an ELISA-like microassay. Standard curves of antibody microarrays are as linear as those of ELISAs. VEGF values were similar to values measured by ELISA. Conclusion In our screen, we found some candidate factors which are currently investigated for their potential of influencing retinopathy after retinal vein occlusion. The use of microarrays to identify protein factors involved in retinal disease in the vitreous will be discussed. [source] | |||||||||||||