Adaptation of Synthetic Peptide Substrate-Based Assays on a Discrete Analyzer

Aiach, Martine; Léon, Martine; Michaud, Anne; Capron, L

doi:10.1055/s-2007-1005022

Cited by 30 publications

(18 citation statements)

References 58 publications

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“…From these intraindividual coefficients of variation, the biological coef ficient of variations can be calculated with the formula CV2to, = CV2assaj, + CV2bioi (ta ble IV) [22], With this biological coefficient of variation it is possible to calculate how much a future result may differ from pre vious values before there is any pathology. There is agreement with several published reports in which normal methods have been adapted to automated instruments [13,15,16,[23][24][25]. Since sample dilution, reagent consistency and precision in reagent han dling are best performed with an automated method, the performance characteristics of this instrument utilizing substrate-based methods are outstanding.…”

Section: Discussionsupporting

confidence: 86%

Rapid Determination of Five Coagulation Parameters in One Sample with a Centrifugal Analyzer

Baś¹,

Costongs²,

Janson³

1987

Pathophysiol Haemos Thromb

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Spectrophotometric clotting assays using chromogenic substrates were adapted for use in a routine laboratory batch analyzer. Contrary to another described method the same plasma dilution can be used for each of the different coagulation parameters. Optimal assay conditions were determined. The within- and between-assay variation, the intraindividual variation during 1 day and from month-to-month were determined; the chromogenic assay was compared to a clotting assay. No significant difference (p < 0.01) between manual and automated methods was found.

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Section: Discussionsupporting

confidence: 86%

Rapid Determination of Five Coagulation Parameters in One Sample with a Centrifugal Analyzer

Baś¹,

Costongs²,

Janson³

1987

Pathophysiol Haemos Thromb

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“…Progressive AT activity was measured using bovine thrombin and S2238 (Biogenic, Mauguio, France) as substrate, as previously described. 21 Immunoreactive AT was measured by means of Laurell immunoelectrophoresis using polyclonal antihuman-AT immunoglobulin G (IgG) antibodies (The Binding Site, Birmingham, United Kingdom). Crossed immunoelectrophoresis was performed with heparin in the first dimension, as described by Sas et al 22 …”

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confidence: 99%

Antithrombin Phe229Leu: a new homozygous variant leading to spontaneous antithrombin polymerization in vivo associated with severe childhood thrombosis

Picard

Dautzenberg

Villoutreix

et al. 2003

Blood

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There is increasing evidence that serpin conformational alteration caused by single point mutations can be responsible for protein deficiency associated with human diseases. A typical example is the 1-antitrypsin deficiency caused by the Z variant carrying a Glu342Lys substitution. Only a few cases of "conforma-tional disease" involving other serpins have been described so far. We investigated a severe antithrombin deficiency in a 13-month-old child with fever and cere-bral venous thrombosis. The infant was found to be homozygous for a new anti-thrombin gene mutation (7396T>C, predicting a Phe229Leu antithrombin variant), and heterozygous for the factor V Leiden mutation. Mild atypical antithrom-bin deficiency was found in both parents, who were first cousins, asymptomatic, and heterozygous for the same antithrom-bin gene mutation. The Phe229Leu variant , which does not readily fit into the current classification of antithrombin deficiency , was shown to be a thermolabile antithrombin that spontaneously polymer-ized in the proband's circulation. This points to a key role for the conserved Phe at position 229, which is near the reactive site loop in a region critical for serpin function and stability. Molecular model-ing suggested how the mutation might destabilize this region of the protein and thereby favor reactive site loop insertion and polymerization. This study provides the first direct evidence of antithrombin polymerization in vivo causing antithrom-bin deficiency and severe thrombotic disease. (Blood. 2003;102:919-925)

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“…AT activity, ie, the total capacity of plasma to neutralize bovine thrombin, was assayed as previously described. 24 Plasma antigen was quantified by electroimmunodiffusion with an immune serum from Diagnostica-Stago. The standard was pooled normal plasma obtained from 30 donors.…”

Section: At Assaysmentioning

confidence: 99%

Three novel mutations of antithrombin inducing high-molecular-mass compounds.

Emmerich

Vidaud

Alhenc‐Gelas

et al. 1994

Arterioscler Thromb

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We have identified three novel mutations of the antithrombin (AT) gene in patients with thrombotic complications: a Cys 128 -» Tyr mutation, a G -» A mutation in the intervening sequence 4 (FVS4) 14 nucleotide 5' to exon 5, and a 9 bp deletion in the 3' end of exon 6 resulting in a short aberrant sequence after Arg 425. The latter mutation was associated with an Arg 47 -• His mutation in two compound heterozygous brothers. These three mutations led to the expression in the circulation of small amounts of inactive molecules with a high molecular mass in immunoblot analysis. In reducing conditions, these variant molecules had a normal molecular mass, which led us to postulate that these mutations prevent the formation of one intramolecular disulfidc bond and allow the formation of intermolecular disulfide bonds. Plasma from a heterozygous patient bearing the Cys 128 -• A ntithrombin (AT) is the main natural inhibitor of /\ thrombin and other coagulation proteases, and X A. its physiological importance was first shown in 1965 by the description of a hereditary AT deficiency in a family with unexplained thrombosis.1 The inhibitory effect of AT, due to the almost irreversible trapping of target proteases, is strongly enhanced by natural glycosaminoglycans such as heparin.The AT gene, the nucleotide sequence of which has recently been elucidated, 2 maps more than 13 480 bp, comprises 7 exons (1, 2, 3a, 3b, 4, 5, and 6), and encodes a polypeptide of 464 amino acids. Several intracellular modifications occur before the secretion of the folded protein, such as the formation of three intramolecular disulfide bonds in positions 8-128, 21-95, and 247-430 of the amino-acid sequence, glycosylation at four sites, and cleavage of a 32-amino-acid signal peptide (for a review see Reference 3). The structure of the mature protein presents large homologies with other serine protease inhibitors, forming a superfamily called serpins. Crystallographic study of cleaved a-1-antitrypsin provided a working model of three-dimensional serpin structure, 4 the validity of which was recently confirmed by crystal

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Adaptation of Synthetic Peptide Substrate-Based Assays on a Discrete Analyzer

Cited by 30 publications

References 58 publications

Rapid Determination of Five Coagulation Parameters in One Sample with a Centrifugal Analyzer

Rapid Determination of Five Coagulation Parameters in One Sample with a Centrifugal Analyzer

Antithrombin Phe229Leu: a new homozygous variant leading to spontaneous antithrombin polymerization in vivo associated with severe childhood thrombosis

Three novel mutations of antithrombin inducing high-molecular-mass compounds.

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