sepharose and Helicobacter-Infections

sepharose has been researched along with Helicobacter-Infections* in 2 studies

Other Studies

2 other study(ies) available for sepharose and Helicobacter-Infections

ArticleYear
Differential effects of DEAE negative mode chromatography and gel-filtration chromatography on the charge status of Helicobacter pylori neutrophil-activating protein.
    PloS one, 2017, Volume: 12, Issue:3

    Helicobacter pylori neutrophil-activating protein (HP-NAP) is involved in H. pylori-associated gastric inflammation. HP-NAP is also a vaccine candidate, a possible drug target, and a potential diagnostic marker for H. pylori-associated diseases. Previously, we purified recombinant HP-NAP by one-step diethylaminoethyl (DEAE) negative mode chromatography by collecting the unbound fraction at pH 8.0 at 4°C. It remains unclear why HP-NAP does not bind to DEAE resins at the pH above its isoelectric point during the purification. To investigate how pH affects the surface net charge of HP-NAP and its binding to DEAE resins during the purification, recombinant HP-NAP expressed in Escherichia coli was subjected to DEAE negative mode chromatography at pH ranging from 7.0 to 9.0 at 25°C and the surface charge of purified HP-NAP was determined by capillary electrophoresis. A minimal amount of HP-NAP was detected in the elution fraction of DEAE Sepharose resin at pH 8.5, whereas recombinant HP-NAP was detected in the elution fraction of DEAE Sephadex resin only at pH 7.0 and 8.0. The purified recombinant HP-NAP obtained from the unbound fractions was not able to bind to DEAE resins at pH 7.0 to 9.0. In addition, the surface charge of the purified HP-NAP was neutral at pH 7.0 to 8.0 and was either neutral or slightly negative at pH 8.5 and 9.0. However, recombinant HP-NAP purified from gel-filtration chromatography was able to bind to DEAE Sepharose resin at pH 7.0 to 9.0 and DEAE Sephadex resin at pH 7.0. At pH 8.5 and 9.0, only the negatively charged species of HP-NAP were found. Thus, recombinant HP-NAP with different charge status can be differentially purified by DEAE negative mode chromatography and gel-filtration chromatography. Furthermore, the charge distribution on the surface of HP-NAP, the presence of impure proteins, and the overall net charge of the resins all affect the binding of HP-NAP to DEAE resins during the negative purification.

    Topics: Bacterial Proteins; Chromatography, Gel; Chromatography, Ion Exchange; DEAE-Dextran; Electrochemistry; Electrophoresis, Capillary; Ethanolamines; Helicobacter Infections; Helicobacter pylori; Humans; Hydrogen-Ion Concentration; Ion Exchange Resins; Neutrophils; Recombinant Proteins; Sepharose

2017
Magnetic immuno-PCR assay with inhibitor removal for direct detection of Helicobacter pylori in human feces.
    Journal of clinical microbiology, 2001, Volume: 39, Issue:10

    A PCR protocol was developed to detect Helicobacter pylori in human stool specimens. This protocol was based on the association of a magnetic immuno-PCR assay with a technique to remove inhibitors (agarose-embedded DNA preparation). Of the 47 H. pylori-positive and 57 H. pylori-negative patients included in this study, 38 were positive and 66 were negative by this new protocol. The sensitivity, specificity, and predictive values for a positive or a negative result were 80.9% (95% confidence interval [CI], 66.3 to 90.4), 100% (95% CI, 92.1 to 100), 100% (95% CI, 88.6 to 100), and 86.4% (95% CI, 75.2 to 93.2), respectively.

    Topics: Adolescent; Adult; Aged; Aged, 80 and over; DNA, Bacterial; Feces; Female; Helicobacter Infections; Helicobacter pylori; Humans; Immunomagnetic Separation; Male; Middle Aged; Polymerase Chain Reaction; Sensitivity and Specificity; Sepharose

2001