Comparison of Glucose Oxidase and Peroxidase as Labels for Antibody in Enzyme-linked Immunosorbent Assay

Johnson, Roy B.; Libby, R; Nakamura, Robert M.

doi:10.1080/01971528008055774

Cited by 16 publications

(10 citation statements)

References 7 publications

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“…Fig. 7 shows the sensorgrams of the interaction between an anti-GH monoclonal antibody (A1-549, Bioclone) and individual GH isoforms (5,17,20, and 22 kDa from recombinant, pituitary and placental origin). From the sensorgrams it is evident that the placental growth hormone variants are not recognised at all (and thus information on the epitope recognised by this antibody is obtained).…”

Section: Cross Reactivity and Secondary Antibodymentioning

confidence: 99%

“…relies on the use of antibodies. At least two antibodies are required to perform a single measurement: one to isolate the hormone of interest from the biological matrix and another one, usually covalently attached to a sensitive read-out device such as horse-radish peroxidise [5], europium [6], acridinium ester [7], etc., for detection purposes. This set-up yields high specificity as usually two monoclonal antibodies are employed.…”

Section: General Aspects Of the Measurements Of Protein Hormonesmentioning

confidence: 99%

See 1 more Smart Citation

Surface plasmon resonance immuno assays – A perspective

Gutiérrez-Gallego

Bosch

Such-Sanmartín

et al. 2009

Growth Hormone & IGF Research

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Section: Cross Reactivity and Secondary Antibodymentioning

confidence: 99%

Section: General Aspects Of the Measurements Of Protein Hormonesmentioning

confidence: 99%

Surface plasmon resonance immuno assays – A perspective

Gutiérrez-Gallego

Bosch

Such-Sanmartín

et al. 2009

Growth Hormone & IGF Research

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“…Periodate method (Nakane's method) (14). HRP solution (15 mg/mL; 1 mL) was added to the periodate solution (0.06 mol/L; 1 mL), stirred gently for 30 min at room temperature and dialysed with bicarbonate solution (0.01 mol/L, pH 9.5) for 12 h at 4°C.…”

Section: Hrp Immobilizationmentioning

confidence: 99%

“…The HRP immobilization methods screened were the adsorption method (12), the glutaraldehyde method (13) and the periodate method (14), and the supports screened were Chitopearl gel and glass beads. The order of activity of the HRP immobilized by the different methods were as follows: Chitopearl/periodate method (15.5 delta absorbance A 505/10 min/100 mg) bChitopearl/adsorption method (4.12) bglass beads/periodate method (1.92) bChitopearl/glutaraldehyde method (1.28) bglass beads/ adsorption method (0.46) bglass beads/glutaraldehyde method (0) (Fig.…”

Section: Immobilization Of Hrp and A¯ow Cell Reactormentioning

confidence: 99%

Determination of hydrogen peroxide by micro-flow injection-chemiluminescence using a coupled flow cell reactor chemiluminometer

Nozaki

Kawamoto

2000

Luminescence

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A novel flow cell reactor was developed for micro-flow injection determination of hydrogen peroxide (H(2)O(2)) using horseradish peroxide (HRP)-catalysed luminol chemiluminescence. The newly developed flow cell reactor for a chemiluminometer allowed mixing of the chemiluminescent reagents in front of a photomultiplier for maximum detection of the emitted light. The rapid mixing allowed a decrease in the flow rate of the pump to 0.1-0.01 mL/min, resulting in increased sensitivity of detection of light. The flow cell reactor was made by packing HRP-immobilized gels into a flow cell (Teflon tube; 6 cm x 0.98 mm i.d.) located in the cell holder of a chemiluminometer (flow-through type). The HRP-immobilized gels were made by immobilizing HRP onto the Chitopearl gel by the periodate method. H(2)O(2) specimens (50 microL) were injected into a stream of water delivered at a flow rate of 0.1 mL/min and mixed with a luminol solution (0.56 mmol/L in Tricine buffer, pH 9.2) delivered at 0.1 mL/min in the flow cell reactor. Within-run reproducibility of the assay of H(2)O(2) was 2.4% (4.85 micromol/L; flow rate 0.1 mL/min, injection interval 10 min). The reproducibility of the H(2)O(2) assay was influenced by the flow rates and the injection intervals of the H(2)O(2) specimens. As the flow rates decreased, both the light intensity and the light duration increased. Optimal light intensity was obtained at a luminol concentration of 3-8 mmol/L, but 0.56 mmol/L was sufficient for assay of H(2)O(2) in clinical specimens. At a luminol concentration of 0.56 mmol/L, the regression equation of the standard curve for H(2)O(2) (0-9.7 micromol/L) was Y = 27.5 X(2) + 394 X + 58.9 (Y = light intensity; X = concentration of H(2)O(2)) and the detection limit of H(2)O(2) was 0.2 micromol/L. This method was used to assay glucose (2.7-16.7 mmol/L) based on a glucose oxidase (20 U/mL, pH 7.4) reaction. The standard curve for glucose was Y = 167 X(2) - 351 X + 1484 (Y = light intensity; X = glucose). The within-run reproducibility for an aqueous glucose standard (2.7 mmol/L) and a control serum (glucose, 5 mmol/L) was 4.48% (n = 5) and 5.70% (n = 9), respectively.

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“…Glucose oxidase-conjugated rabbit anti-chinchilla immunoglobulin and the buffered sub¬ strate were prepared according to the methods of Johnson et al 12 All diluent was PBS containing 0.05% v/v polysorbate 20. A 60-mL aliquot of the bacterial suspension (antigen) was added to each of the 96 flat-bottomed wells of a polystyrene plate (Immulon I).…”

Section: Bacteriamentioning

confidence: 99%

Antibody Response in Experimental Haemophilus influenzae Otitis Media

Yamaguchi

DeMaria

Lim³

1986

Archives of Otolaryngology - Head and Neck Surgery

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\s=b\Because Haemophilus influenzae is one of the most common pathogens in otitis media with effusion, we have investigated the antibody response in the serum and middle ear effusion (MEE) in nontypable H influenzae\p=n-\inducedexperimental acute otitis media in the chinchilla, using an enzyme-linked immunosorbent assay. During acute otitis media, a reasonable antibody titer was observed: local antibody titers in the MEEs were equal to or greater than those of the serum samples for four of five MEE samples obtained at one week after inoculation. By two weeks, titers in both serum and MEE samples were increased significantly. However, the bacteria persisted in the middle ear cavity, even in the presence of increasing antibody titers, for up to five weeks after infection. The reason for this persistence of the bacteria in the middle ear is unknown. (Arch Otolaryngol Head Neck Surg 1986;112:554-557) TJaemophilus influenzae, in partic--L Jular the nontypable strains, is one of the most common bacteria iso¬ lated from the middle ear effusions (MEEs) of patients with acute or chronic otitis media with effusion (OME).1"3 Our laboratory recently re¬ ported that the nontypable biotype II strain is the most common pathogen associated with cases of chronic OME4 and that nonviable H influenzae or endotoxin from this organism may also be responsible for the induction of OME.56 There is reason to believe that the endotoxin, a cell wall compo¬ nent of gram-negative bacteria, may be responsible for inducing MEEs and pathologic changes in the middle ear, because a large number of MEE sam¬ ples from patients with chronic OME contained an appreciable amount of endotoxin even when the presence of bacteria could not be demonstrated by culture or Gram's stain.7Even though the underlying reason for the biotype II strain's prevalence in chronic OME is poorly understood, one factor could be the poor immunogenicity of this organism. Little infor¬ mation is available regarding the sys¬ temic and local immune responses against nontypable H influenzae dur¬ ing middle ear infection. Work in this area has been hampered because these bacteria appear to be antigenically heterogeneous,8 and the antigens of this organism that confer protection to the host are undefined.9 The pur¬ pose of this study was to investigate the fundamentals of the antibody responses in serum and MEE samples in the chinchilla after challenge with viable nontypable H influenzae (biotype II). MATERIALS AND METHODS BacteriaThe H influenzae nontypable isolate (serologically not typable, but belonging to biotype II), originally isolated from a case of otitis media, was obtained from the Ohio Department of Health Laboratories, Columbus. The bacteria were cultured and used as the inoculum, as described previously.-Experimental Design Forty-one healthy chinchillas (310 to 670 g) without evidence of previous or current infection were used in this study. Thirtyfour chinchillas were aseptically inocu¬ lated directly into the left superior bulla with 0.3 mL of bacterial suspensio...

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Comparison of Glucose Oxidase and Peroxidase as Labels for Antibody in Enzyme-linked Immunosorbent Assay

Cited by 16 publications

References 7 publications

Surface plasmon resonance immuno assays – A perspective

Surface plasmon resonance immuno assays – A perspective

Determination of hydrogen peroxide by micro-flow injection-chemiluminescence using a coupled flow cell reactor chemiluminometer

Antibody Response in Experimental Haemophilus influenzae Otitis Media

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