Bacterial Antibodies

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Dongqing Li - One of the best experts on this subject based on the ideXlab platform.

  • multiplexed high throughput electrokinetically controlled immunoassay for the detection of specific Bacterial Antibodies in human serum
    Analytica Chimica Acta, 2008
    Co-Authors: Philip M Sherman, Dongqing Li
    Abstract:

    Abstract In previous studies we have developed a simple electrokinetically-controlled lab-on-a-chip for heterogeneous immunoassay. In that method, all the sequential operations in an immunoassay, such as reagent loading and washing, were performed automatically by electrokinetically controlling the flow in an H-shaped microchannel. Here, we demonstrated further development of a high-throughput immunoassay microfluidic chip, and the application of the new immunoassay microfluidic chip in assaying human serum. The microfluidic immunoassay analyzed ten samples in parallel in 22 min. Bacterial Antibodies in samples were captured by antigens pre-patterned on the bottom wall of a microchannel and then bound with TRITC-labeled detection Antibodies to generate fluorescent signals. With optimized surface concentration of antigen, the assay detected Escherichia coli O157:H7 antibody and Helicobacter pylori antibody from buffer solutions in concentration ranges of 0.02–10 μg mL −1 and 0.1–50 μg mL −1 , respectively. Human sera that were E. coli -positive or H. pylori -positive were accurately distinguished from respective negative controls. Moreover, the two Antibodies, anti- E. coli and anti- H. pylori Antibodies, could be simultaneously detected from human serum. This electrokinetically-controlled immunoassay shows an excellent potential for efficiently detecting multiple pathogenic infections in clinical environments.

  • Multiplexed High-Throughput Electrokinetically-Controlled Immunoassay on a Chip for the Detection of Specific Bacterial Antibodies in Human Serum
    Volume 11: Micro and Nano Systems Parts A and B, 2007
    Co-Authors: Philip M Sherman, Dongqing Li
    Abstract:

    This work presents a multiplexed electrokinetically-controlled heterogeneous immunoassay that can process ten samples in parallel. The immunoassay microchip was soft-lithographically fabricated using poly(dimethylsiloxane) and glass. Controlling parameters of the electrokinetically-driven flow in the microfluidic network was determined by numerically simulating transport processes. Multiple passively adsorbed antigens captured Antibodies present in samples, which then bound with TRITC-labeled detection Antibodies to generate fluorescent signals. Antibodies against Escherichia coli O157:H7 and Helicobacter pylori were studied as model analytes. After conditions for antigen-coating were optimized, a 24-minute assay detected E. coli O157:H7 antibody in the concentration range of 0.02–10 μg/mL, and H. pylori antibody in the range of 0.1–50 μg/mL. In testing human serum samples, non-specific binding of serum components was effectively suppressed by using 10% (w/v) bovine serum albumin. An accuracy of 100% was achieved in detecting either E. coli O157:H7 antibody or H. pylori antibody from human serum samples. Simultaneous screening of both Antibodies was also successfully demonstrated. The immunoassay chip shows an excellent potential for efficiently detecting multiple pathogenic infections in clinical environments.Copyright © 2007 by ASME

Isao Okayasu - One of the best experts on this subject based on the ideXlab platform.

  • fusobacterium varium localized in the colonic mucosa of patients with ulcerative colitis stimulates species specific antibody
    Journal of Gastroenterology and Hepatology, 2002
    Co-Authors: Toshifumi Ohkusa, Nobuhiro Sato, Tatuo Ogihara, Koji Morita, Masayuki Ogawa, Isao Okayasu
    Abstract:

    Abstract Background: Microbial agents are a possible cause of ulcerative colitis. We have previously reported evidence of bacteria invading the colonic mucosa of patients with ulcerative colitis. We have isolated bacteria from inflamed colonic mucosa, examined the localization of the species in the mucosa, and assayed for serum Antibodies to the bacteria. Methods: Cohorts of 31 per group were enrolled from patients with active ulcerative colitis, Crohn's disease, ischemic colitis, and colon adenomas. A group of 31 healthy controls were also studied. The presence of bacteria in biopsies of patients with ulcerative colitis was analyzed by both isolation and immunohistochemistry. Sera from patients were tested for Bacterial Antibodies using both Western blots and enzyme-linked immunosorbent assay (ELISA). Results: Only sera from patients with ulcerative colitis gave specific reactions with Fusobacterium varium in Western blot assays. The detection rate of specific bands was higher for patients with ulcerative colitis (61%) than for subjects with either Crohn's disease (13%) or healthy controls (29%) (P < 0.001 and P = 0.021, respectively). The ELISA showed that the mean optical densities with extracts of F. varium as antigen were significantly higher for ulcerative colitis patients than for subjects with either Crohn's disease or healthy controls (P < 0.001). Immunohistochemical detection of F. varium in colonic mucosa was significantly higher in patients with ulcerative colitis (84%) than for subjects with either Crohn's disease (16%) or other controls (3–13%) (P < 0.001). Conclusions:Fusobacterium varium bacteria were present in a significant number of patients with active ulcerative colitis, and should be tested in therapeutic trials in order to confirm the causal relationship between F. varium and ulcerative colitis.

Philip M Sherman - One of the best experts on this subject based on the ideXlab platform.

  • multiplexed high throughput electrokinetically controlled immunoassay for the detection of specific Bacterial Antibodies in human serum
    Analytica Chimica Acta, 2008
    Co-Authors: Philip M Sherman, Dongqing Li
    Abstract:

    Abstract In previous studies we have developed a simple electrokinetically-controlled lab-on-a-chip for heterogeneous immunoassay. In that method, all the sequential operations in an immunoassay, such as reagent loading and washing, were performed automatically by electrokinetically controlling the flow in an H-shaped microchannel. Here, we demonstrated further development of a high-throughput immunoassay microfluidic chip, and the application of the new immunoassay microfluidic chip in assaying human serum. The microfluidic immunoassay analyzed ten samples in parallel in 22 min. Bacterial Antibodies in samples were captured by antigens pre-patterned on the bottom wall of a microchannel and then bound with TRITC-labeled detection Antibodies to generate fluorescent signals. With optimized surface concentration of antigen, the assay detected Escherichia coli O157:H7 antibody and Helicobacter pylori antibody from buffer solutions in concentration ranges of 0.02–10 μg mL −1 and 0.1–50 μg mL −1 , respectively. Human sera that were E. coli -positive or H. pylori -positive were accurately distinguished from respective negative controls. Moreover, the two Antibodies, anti- E. coli and anti- H. pylori Antibodies, could be simultaneously detected from human serum. This electrokinetically-controlled immunoassay shows an excellent potential for efficiently detecting multiple pathogenic infections in clinical environments.

  • Multiplexed High-Throughput Electrokinetically-Controlled Immunoassay on a Chip for the Detection of Specific Bacterial Antibodies in Human Serum
    Volume 11: Micro and Nano Systems Parts A and B, 2007
    Co-Authors: Philip M Sherman, Dongqing Li
    Abstract:

    This work presents a multiplexed electrokinetically-controlled heterogeneous immunoassay that can process ten samples in parallel. The immunoassay microchip was soft-lithographically fabricated using poly(dimethylsiloxane) and glass. Controlling parameters of the electrokinetically-driven flow in the microfluidic network was determined by numerically simulating transport processes. Multiple passively adsorbed antigens captured Antibodies present in samples, which then bound with TRITC-labeled detection Antibodies to generate fluorescent signals. Antibodies against Escherichia coli O157:H7 and Helicobacter pylori were studied as model analytes. After conditions for antigen-coating were optimized, a 24-minute assay detected E. coli O157:H7 antibody in the concentration range of 0.02–10 μg/mL, and H. pylori antibody in the range of 0.1–50 μg/mL. In testing human serum samples, non-specific binding of serum components was effectively suppressed by using 10% (w/v) bovine serum albumin. An accuracy of 100% was achieved in detecting either E. coli O157:H7 antibody or H. pylori antibody from human serum samples. Simultaneous screening of both Antibodies was also successfully demonstrated. The immunoassay chip shows an excellent potential for efficiently detecting multiple pathogenic infections in clinical environments.Copyright © 2007 by ASME

Toshifumi Ohkusa - One of the best experts on this subject based on the ideXlab platform.

  • fusobacterium varium localized in the colonic mucosa of patients with ulcerative colitis stimulates species specific antibody
    Journal of Gastroenterology and Hepatology, 2002
    Co-Authors: Toshifumi Ohkusa, Nobuhiro Sato, Tatuo Ogihara, Koji Morita, Masayuki Ogawa, Isao Okayasu
    Abstract:

    Abstract Background: Microbial agents are a possible cause of ulcerative colitis. We have previously reported evidence of bacteria invading the colonic mucosa of patients with ulcerative colitis. We have isolated bacteria from inflamed colonic mucosa, examined the localization of the species in the mucosa, and assayed for serum Antibodies to the bacteria. Methods: Cohorts of 31 per group were enrolled from patients with active ulcerative colitis, Crohn's disease, ischemic colitis, and colon adenomas. A group of 31 healthy controls were also studied. The presence of bacteria in biopsies of patients with ulcerative colitis was analyzed by both isolation and immunohistochemistry. Sera from patients were tested for Bacterial Antibodies using both Western blots and enzyme-linked immunosorbent assay (ELISA). Results: Only sera from patients with ulcerative colitis gave specific reactions with Fusobacterium varium in Western blot assays. The detection rate of specific bands was higher for patients with ulcerative colitis (61%) than for subjects with either Crohn's disease (13%) or healthy controls (29%) (P < 0.001 and P = 0.021, respectively). The ELISA showed that the mean optical densities with extracts of F. varium as antigen were significantly higher for ulcerative colitis patients than for subjects with either Crohn's disease or healthy controls (P < 0.001). Immunohistochemical detection of F. varium in colonic mucosa was significantly higher in patients with ulcerative colitis (84%) than for subjects with either Crohn's disease (16%) or other controls (3–13%) (P < 0.001). Conclusions:Fusobacterium varium bacteria were present in a significant number of patients with active ulcerative colitis, and should be tested in therapeutic trials in order to confirm the causal relationship between F. varium and ulcerative colitis.

Valeria Szijarto - One of the best experts on this subject based on the ideXlab platform.

  • anti Bacterial monoclonal Antibodies
    Advances in Experimental Medicine and Biology, 2017
    Co-Authors: Eszter Nagy, Gabor Nagy, Christine Power, Adriana Badarau, Valeria Szijarto
    Abstract:

    The failing efficacy of antibiotics and the high mortality rate among high-risk patients calls for new treatment modalities for Bacterial infections. Due to the vastly divergent pathogenesis of human pathogens, each microbe requires a tailored approach. The main modes of action of anti-Bacterial Antibodies are virulence factor neutralization, complement-mediated Bacterial lysis and enhancement of opsonophagocytic uptake and killing (OPK). Gram-positive bacteria cannot be lysed by complement and their pathogenesis often involves secreted toxins, therefore typically toxin-neutralization and OPK activity are required to prevent and ameliorate disease. In fact, the success stories in terms of approved products, in the anti-Bacterial mAb field are based on toxin neutralization (Bacillus anthracis, Clostridium difficile). In contrast, Gram-negative bacteria are vulnerable to antibody-dependent complement-mediated lysis, while their pathogenesis rarely relies on secreted exotoxins, and involves the pro-inflammatory endotoxin (lipopolysaccharide). Given the complexity of Bacterial pathogenesis, antibody therapeutics are expected to be most efficient upon targeting more than one virulence factor and/or combining different modes of action. The improved understanding of Bacterial pathogenesis combined with the versatility and maturity of antibody discovery technologies available today are pivotal for the design of novel anti-Bacterial therapeutics. The intensified research generating promising proof-of-concept data, and the increasing number of clinical programs with anti-Bacterial mAbs, indicate that the field is ready to fulfill its promise in the coming years.