New Rapid Enzyme-Linked Immunosorbent Assay to Detect Antibodies against Bacterial Surface Antigens Using Filtration Plates.

Itoh, Shinji; Kariya, Masako; Nagano, Keiji; Yokoyama, Shin-ichiro; Fukao, Toshio; Yamazaki, Yoko; Mori, Hiroshi

doi:10.1248/bpb.25.986

Cited by 27 publications

(21 citation statements)

References 24 publications

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“…To quantitatively detect the presence of extracellular CsgA as high-molecular-weight assemblies, an adapted whole-cell filtration ELISA protocol for detecting bacterial surface antigens was used 55 . BIND transformants were used to inoculate 3 ml YESCA liquid cultures supplemented with 50 mg ml À 1 of carbenicillin, grown to mid-log phase and induced with 0.25 mM of IPTG.…”

Section: And 3)mentioning

confidence: 99%

“…To a Multiscreen-GV 96-well filter plate (0.22 mm pore size; EMD Millipore), 25 ml of the diluted culture was added and washed three times in wash buffer (TBS þ 0.1% Tween-20 þ 0.1% NaN 3 ) and incubated in 200 ml of Blocking Buffer (wash buffer supplemented with 1% bovine serum albumin and 0.01% H 2 O 2 ) for 1 h at 37°C. The H 2 O 2 is needed to inactivate endogenous cellular peroxidases 55 . The samples were then washed three times in wash buffer, incubated with anti-CsgA primary antibody 8 diluted to 1:10,000 in wash buffer for 1 h at 25°C, washed three more times in wash buffer and then incubated with goat anti-rabbit HRP-conjugated secondary antibody (Thermo Fisher Scientific) diluted to 1:5,000 in wash buffer for 1 h at 25°C.…”

Section: And 3)mentioning

confidence: 99%

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Programmable biofilm-based materials from engineered curli nanofibres

et al. 2014

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The significant role of biofilms in pathogenicity has spurred research into preventing their formation and promoting their disruption, resulting in overlooked opportunities to develop biofilms as a synthetic biological platform for self-assembling functional materials. Here we present Biofilm-Integrated Nanofiber Display (BIND) as a strategy for the molecular programming of the bacterial extracellular matrix material by genetically appending peptide domains to the amyloid protein CsgA, the dominant proteinaceous component in Escherichia coli biofilms. These engineered CsgA fusion proteins are successfully secreted and extracellularly self-assemble into amyloid nanofibre networks that retain the functions of the displayed peptide domains. We show the use of BIND to confer diverse artificial functions to the biofilm matrix, such as nanoparticle biotemplating, substrate adhesion, covalent immobilization of proteins or a combination thereof. BIND is a versatile nanobiotechnological platform for developing robust materials with programmable functions, demonstrating the potential of utilizing biofilms as large-scale designable biomaterials.

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Section: And 3)mentioning

confidence: 99%

Section: And 3)mentioning

confidence: 99%

Programmable biofilm-based materials from engineered curli nanofibres

et al. 2014

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“…Filtration ELISA to Detect Antibodies against E. coli Whole Cells The procedures for the pre-absorption of samples with C600 and filtration ELISA were essentially the same as described previously. 17) Washing buffer was PBS containing 0.1% Tween 20. Blocking buffer-A consisted of PBS containing 1% bovine serum albumin (Fraction V, Nacalai Tesque Inc., Kyoto, Japan), 5% normal calf serum (Irvin Scientific, Kogan, UT, U.S.A.), 0.1% Tween 20 and 0.1% NaN 3 .…”

Section: Detection Of Fecal Bacteriamentioning

confidence: 99%

“…We also reported that fecal immunoglobulin A (IgA) antibodies against EHEC O157:H7 increased not only in children infected with the pathogen 17) but also in mice inoculated orally. 18) Camara et al 19) demonstrated that colostrum IgA antibodies against intimin of enteropathogenic E. coli, which have a LEE cluster similar to that of EHEC, inhibited the adherence of the bacteria to HeLa cells.…”

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Adhesion of Enterohemorrhagic <i>Escherichia coli</i> O157:H7 to the Intestinal Epithelia Is Essential for Inducing Secretory IgA Antibody Production in the Intestine of Mice

Nagano

Taguchi

Tokoro

et al. 2014

Biological & Pharmaceutical Bulletin

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We examined whether adherence of enterohemorrhagic Escherichia coli (EHEC) O157:H7 to intestinal epithelial cells contributed to the induction of secretory immunoglobulin A (IgA) antibody production in mice. Wild-type EHEC O157:H7 and its mutants deficient in the espA, sepL, tir and eae genes, encoding adherent factors on the locus of enterocyte effacement (LEE), were inoculated intragastrically into mice. Inoculation of wild-type EHEC induced fecal IgA antibodies specific to EHEC at 4 weeks after the inoculation, but that of espA-and sepL-deletion mutants did not. Furthermore, even 4 inoculations at weekly intervals with espA-deletion mutant, heat-killed wild-type EHEC and nonpathogenic E. coli did not induce fecal IgA antibodies, although these bacterial inoculations induced serum antibodies. Kanamycin (Km)-treated mice showed prolonged and similar fecal shedding of Km-resistant mutants of EHEC O157:H7 including A2-F6 having intact LEE, A6-E7 (sepL-insertion mutant), G1-E11 (tir-insertion mutant) and Δeae (eae-deletion mutant). In this case, A2-F6 induced fecal IgA antibodies, but the other mutants with defective LEE did not. In contrast to the fecal IgA antibodies, serum IgM and IgG antibodies were induced in mice inoculated with any of the LEE defective mutants as well as A2-F6. Thus, adhesion of EHEC to epithelial cells is essential for inducing the mucosal immune response in the intestine but not for inducing the systemic immune response.

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“…Itoh et al used the 96-well filtration plate to concentrate the microorganisms from the sample and detected them by indirect enzymelinked immune sorbent assay (indirect-ELISA) (Itoh et al, 2002). Sardinha et al immobilized the capture antibody on filter membrane to detect pesticide (Sardinha et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

Development of a novel bead-based 96-well filtration plate competitive immunoassay for the detection of Gentamycin

Chan

et al. 2013

Biosensors and Bioelectronics

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New Rapid Enzyme-Linked Immunosorbent Assay to Detect Antibodies against Bacterial Surface Antigens Using Filtration Plates.

Cited by 27 publications

References 24 publications

Programmable biofilm-based materials from engineered curli nanofibres

Programmable biofilm-based materials from engineered curli nanofibres

Adhesion of Enterohemorrhagic <i>Escherichia coli</i> O157:H7 to the Intestinal Epithelia Is Essential for Inducing Secretory IgA Antibody Production in the Intestine of Mice

Development of a novel bead-based 96-well filtration plate competitive immunoassay for the detection of Gentamycin

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