Effect of surfactants on the intensity of chemiluminescence emission from acridinium ester labelled proteins

Bagazgoitia, F. Javier; García, Jorge Mendoza; Diéquez, Carlos; Weeks, Ian; Woodhead, J. S.

doi:10.1002/bio.1170020303

Cited by 29 publications

(21 citation statements)

References 8 publications

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“…O 2 Àinduced lucigenin chemiluminescence was strongly amplified by Triton X-100. Similarly, Bagazgoitia et al (8) reported that detergents enhanced the CL emission of acridinium esters. A possible explanation could be that the detergent molecules increase the half-life of the superoxide anion species by trapping them so that the superoxide species cannot react with one another by dismutation.…”

Section: Discussionmentioning

confidence: 88%

Enhanced sensitivity of Cypridina luciferin analogue (CLA) chemiluminescence for the detection of .O2 with non‐ionic detergents

Osman¹,

Laane²,

Hilhorst³

2001

Luminescence

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Superoxide anion-triggered chemiluminescence of Cypridina luciferin analogue (CLA), 2-methyl-6-phenyl-3,7-dohydroimidazo[1,2-a]pyrazin-3-one, is enhanced by non-ionic detergents such as Tween 20, Triton X-100 and Tween 80. At the concentration of 0.6% (v/v) the largest increase (2.7-fold) of CLA light intensity was obtained with Tween 20, followed by Tween 80 and Triton X-100. Using this detergent-amplified CLA chemiluminescence, the detection limits of xanthine and xanthine oxidase were examined at pH 7.4 and reinvestigated at pH 5.5. At pH 5.5, concentrations of xanthine and xanthine oxidase as low as 5 nmol/L and 3.85 Â 10 À7 U/mL, respectively, could be accurately determined, whereas, under the experimental conditions used, at pH 7.4 the lowest concentrations of xanthine and xanthine oxidase detectable were 10 nmol/L and 3.85 Â 10 À6 U/mL. The lowest detectable values of xanthine and xanthine oxidase obtained at pH 5.5 are about 400 and 10 times lower than those previously reported. The detection limit of xanthine (5 nmol/L) by this chemiluminescent-based method is about 200 and 20 times more sensitive than the determination of xanthine by enzymatic means or by HPLC with detection limits of 1 mmol/L and 0.1 mmol/L, respectively. Our data suggest that this chemiluminescent probe can detect concentrations of superoxide anion below the nanomolar range.

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Section: Discussionmentioning

confidence: 88%

Enhanced sensitivity of Cypridina luciferin analogue (CLA) chemiluminescence for the detection of .O2 with non‐ionic detergents

Osman¹,

Laane²,

Hilhorst³

2001

Luminescence

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“…Acridinium salts are known to generate light in the presence of alkaline hydrogen peroxide (1,2). However, many of these acridinium salts are unstable in aqueous solutions and form an equilibrium with their pseudobases (3). These pseudo-bases react very slowly with hydrogen peroxide and consequently the efficiency of the chemiluminescent reaction is strongly reduced (3).…”

Section: Introductionmentioning

confidence: 99%

“…However, many of these acridinium salts are unstable in aqueous solutions and form an equilibrium with their pseudobases (3). These pseudo-bases react very slowly with hydrogen peroxide and consequently the efficiency of the chemiluminescent reaction is strongly reduced (3). To circumvent this serious practical limitation, a new class of synthetic acridinium derivatives, which act as substrates for peroxidase enzymes, have recently been introduced (4).…”

Section: Introductionmentioning

confidence: 99%

Comparative studies of the chemiluminescent horseradish peroxidase-catalysed peroxidation of acridan (GZ-11) and luminol reactions: effect of pH and scavengers of reactive oxygen species on the light intensity of these systems

et al. 2000

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In this study, the chemiluminescent horseradish peroxidase/H(2)O(2)-catalysed oxidation of acridan (GZ-11) substrate was compared with the well-characterized light-producing luminol reaction. p-Iodophenol and p-phenylphenol were used as enhancers, respectively, for the luminol and acridan reactions. These two light-producing systems showed significant differences in relation to the effect of pH, as well as the effect of scavengers of reactive oxygen species, on the light intensity. Light production measured could be as low as pH 2.6 in the acridan reaction, whereas light emission was not detected in the luminol system below pH 5.6. In contrast with the luminol system, it was found that superoxide dismutase does not inhibit the light intensity of the acridan system. This suggests that superoxide anion does not participate in the mechanism of the light-emitting steps of the acridan reaction. Addition of hydroxyl radical scavengers, mannitol and benzoate, to the acridan reaction medium had no appreciable effect on the chemiluminescent intensity, indicating that hydroxyl radicals do not interfere in light-emitting steps. In addition, the peroxidation of the acridan substrate was found to be very slow at pH 5.6 in the absence of the enhancer, p-phenylphenol, whereas in its presence a rapid degradation of the acridan substrate was observed. Therefore, it is suggested that the enhancer might be initially oxidized by the HRP/H(2)O(2) system, resulting in the formation of the enhancer radical, which could be the actual oxidizing agent of the acridan substrate. Together, the data presented in this paper indicate that the chemiluminescent horseradish peroxidase-catalysed peroxidation of acridan (GZ-11) is more specific than the luminol reaction for the reactive oxygen species involved in the light-emitting steps, i. e, H(2)O(2).

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“…Firstly, the succinimidyl aryl acridinium ester-labelled proteins are easily prepared in large quantities and show very high stability. Secondly, the speed of quantitation is very rapid; a 6 s integration time has been employed here but the speed of the CL reaction is such that integration times of 2 s or less can be used, mostly when surfactants as CTAC are used to improve the CL reaction (Bagazgoitia et al, 1988).…”

Section: Discussionmentioning

confidence: 99%

“…Optimal conditions of CL measurement, the effect of surfactants on CL-reaction and assay buffer constituents used to avoid non-specific adsorption of analyte to the tube wall, were reported by us in a previous work (Bagazgoitia et al, 1988).…”

Section: Luminometrymentioning

confidence: 95%

Chemiluminescent immunoassay (CLIA) for urinary albumin

Bagazgoitia¹,

García²,

Ibarra³

et al. 1989

J. Biolumin. Chemilumin.

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A simple chemiluminescent immunoassay (CLIA) for urinary albumin has been developed based on the use of a chemiluminescent acridinium ester-labelled human albumin and a commercially available antiserum. It includes two incubation steps and a second polyethylene glycol-assisted antibody separation. The sensitivity of detection is 0.016 mg/l, the assay working range is 0.1-5 mg/l, and the inter-assay CVs are less than or equal to 15%. Using 10- and 50-fold sample dilutions in assay buffer, a wide working range (1-250 mg/l) is obtained covering normal and pathological conditions. Timed overnight urine samples (bed rest conditions) were collected on three consecutive days for each patient. Albumin excretion rate (AER) was 4.7 +/- 2.7 micrograms/min (mean +/- SD), range 1-15.9 micrograms/min in 36 healthy subjects (17 male, 19 female, ages 4-56 years), with day-to-day variations of 28.5 +/- 20% (mean +/- SD), range 3.3-76.1%. The use of an acridinium ester as a chemiluminescent (CL) label overcomes the disadvantages of short shelf-life and health and safety hazards associated with radioisotopes. Results compare favourably with those obtained using a commercially available RIA kit.

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Effect of surfactants on the intensity of chemiluminescence emission from acridinium ester labelled proteins

Cited by 29 publications

References 8 publications

Enhanced sensitivity of Cypridina luciferin analogue (CLA) chemiluminescence for the detection of .O2 with non‐ionic detergents

Enhanced sensitivity of Cypridina luciferin analogue (CLA) chemiluminescence for the detection of .O2 with non‐ionic detergents

Comparative studies of the chemiluminescent horseradish peroxidase-catalysed peroxidation of acridan (GZ-11) and luminol reactions: effect of pH and scavengers of reactive oxygen species on the light intensity of these systems

Chemiluminescent immunoassay (CLIA) for urinary albumin

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