Fast double antibody radioimmunoassay of human granolocyte myeloperoxidase and its application to plasma

Pincemaïl, Joël; Deby‐Dupont, G.; Deby, C.; Thirion, Anne-Marie; Torpier, Gérard; Faymonville, Marie-Élisabeth; Damas, Pierre; Tomassini, Margherita; Lamy, Maurice; Franchimont, P

doi:10.1016/0022-1759(91)90023-9

Cited by 25 publications

(18 citation statements)

References 18 publications

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“…Plasma concentrations of TNF-a were measured with an immunoenzymatic assay kit (Medgenic Diagnostics, Fleurus, Belgium). Plasma MPO concentration was assessed according to the radioimmunological method described by Pincemail et al (1991).…”

Section: Blood Samplingmentioning

confidence: 99%

Endotoxaemia, production of tumour necrosis factor ? and polymorphonuclear neutrophil activation following strenuous exercise in humans

Camus

Nys

Poortmans

et al. 1998

European Journal of Applied Physiology

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To examine whether endotoxaemia accompanying long-term, strenuous physical exercise is involved in exercise-induced increase in plasma tumour necrosis factor alpha (TNF-alpha) concentration and polymorphonuclear neutrophil (PMN) activation, 14 male recreational athletes [mean age 28 (SEM 1) years] were studied. Exercise consisted of a 1.5-km river swim, a 40-km bicycle race, and a 10-km road race. Mean time to complete the race was 149.8 (SEM 4.8) min. The plasma concentrations of granulocyte myeloperoxidase (MPO) and TNF-alpha were significantly higher than baseline values immediately and 1 h after exercise (P<0.001). Both variables returned to pre-race levels the day after exercise. Marked, transient decreases in plasma concentrations of anti-lipopolysaccharide (LPS) immunoglobulin G (IgG) and immunoglobulin M (IgM) antibodies directed against a panel of selected smooth gram-negative LPS were observed after the race, reaching in most cases minimal values in the blood sample drawn immediately following the completion of the triathlon. There was no significant correlation between the magnitude of PMN activation, as assessed by the increase in plasma concentrations of MPO, and the humoral markers of endotoxaemia and TNF-alpha. An inverse, highly significant relationship between the increase in plasma TNF-alpha concentrations and the changes in circulating anti-LPS IgM antibodies concentrations was observed (r = -0.7; P<0.01). These findings suggest that exercise-induced endotoxaemia was involved in the release of TNF-alpha, that the magnitude of the TNF-alpha response to exercise was down-regulated by anti-LPS antibodies of the IgM class, and that the production of TNF-alpha and endotoxaemia did not seem to play a role in the activation of circulating PMN in the exercising subjects.

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Section: Blood Samplingmentioning

confidence: 99%

Endotoxaemia, production of tumour necrosis factor ? and polymorphonuclear neutrophil activation following strenuous exercise in humans

Camus

Nys

Poortmans

et al. 1998

European Journal of Applied Physiology

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“…MPO was purified from human neutrophils by detergent enhanced extraction followed by two steps of chromatography, and antisera were obtained by rabbit immunization against the pure human MPO emulsified with complete Freud adjuvant as previously described [9,21]. Polyclonal antibody (IgG), purified from rabbit antisera by affinity chromatography on Protein A sepharose (GE Healthcare, The Netherlands), was characterized for its specificity and stability and firstly used to develop a radioimmunoassay for total MPO assay in human plasma [9].…”

Section: Purification Of Human Mpo and Rabbit Anti-human Mpo Polyclonmentioning

confidence: 99%

“…Polyclonal antibody (IgG), purified from rabbit antisera by affinity chromatography on Protein A sepharose (GE Healthcare, The Netherlands), was characterized for its specificity and stability and firstly used to develop a radioimmunoassay for total MPO assay in human plasma [9].…”

Section: Purification Of Human Mpo and Rabbit Anti-human Mpo Polyclonmentioning

confidence: 99%

“…The cross-reactivity of the anti-MPO antibodies with albumin, haemoglobin, elastase or eosinophil peroxidase was measured as previously described [9]. The detection limit was defined as the lowest active MPO concentration (mean value ± SD) that was statistically different from the value obtained with buffer alone.…”

Section: Validation Of the Siefed Techniquementioning

confidence: 99%

“…In these conditions, MPO is able to exert a deleterious oxidant activity on neighbouring cells and tissues, and a therapeutic target should be to modulate its activity. Increased blood levels of MPO are now considered as markers of neutrophil activation and degranulation [7,8] and are measured by immunological methods (ELISA or RIA), which quantify the total concentration of the enzyme [9,10] without reflecting its true enzymatic activity. The activity of MPO is currently measured by the direct addition in the medium of H 2 O 2 and a chromogenic or fluorogenic substrate as electron donors [11][12][13].…”

Section: Introductionmentioning

confidence: 99%

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A new easy method for specific measurement of active myeloperoxidase in human biological fluids and tissue extracts

Franck

Kohnen

Boudjeltia³

et al. 2009

Talanta

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a b s t r a c tThe SIEFED ("Specific Immunological Extraction Followed by Enzymatic Detection") method already developed for the specific detection of the activity of equine myeloperoxidase (MPO) was adapted for the specific measurement of active human MPO in biological fluids or tissue extracts. The method consists of the extraction of MPO from aqueous solutions by immobilized anti-MPO antibodies followed by a washing (to eliminate the extraction medium and the biological fluid with their possible interfering molecules) and the measurement of the activity of MPO with a detection system containing a fluorogenic substrate, H 2 O 2 and nitrite ions as reaction enhancer. The SIEFED was applied to study active MPO in human biological fluids (plasma, bronchoalveolar lavage fluid and supernatant from carotids extracts). The SIEFED for human MPO has a sensitivity limit of 0.080 mU/mL and showed good precision with intra-and inter-assay coefficients of variation below 10 and 20% respectively within a broad range of MPO activities establish from 0.156 to 473 mU/mL. The SIEFED for human MPO will be useful for the specific detection of active MPO in complex fluids and can be complementary to an ELISA to determine an active/total MPO ratio in healthy volunteers and patients especially in case of chronic or acute inflammatory diseases.

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Peroxidase‐catalysed oxidation of different dibenzazepine derivatives

Liégeois

Rogister

Delarge

et al. 1995

Archiv der Pharmazie

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According to a recent hypothesis suggesting the potential role of free radical formation in the clozapine-induced agranulocytosis, we have evaluated the susceptibility to the peroxidase-mediated oxidation of different dibenzazepine analogues. On the one hand, compounds with an arylamine group such as clozapine or isoclozapine present a high reactivity in the horseradish peroxidase or myeloperoxidase systems and, on the other hand, fluperlapine, though known to induce agranulocytosis, and other dibenzothiazepine and dibenzoxazepine derivatives appear insensitive to oxidation. Consequently, among tricyclic derivatives, the way of diaryloxa- and diarylthiazepine compounds could be an alternative for the development of safer drugs such as antipsychotics.

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Fast double antibody radioimmunoassay of human granolocyte myeloperoxidase and its application to plasma

Cited by 25 publications

References 18 publications

Endotoxaemia, production of tumour necrosis factor ? and polymorphonuclear neutrophil activation following strenuous exercise in humans

Endotoxaemia, production of tumour necrosis factor ? and polymorphonuclear neutrophil activation following strenuous exercise in humans

A new easy method for specific measurement of active myeloperoxidase in human biological fluids and tissue extracts

Peroxidase‐catalysed oxidation of different dibenzazepine derivatives

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