Purification of bovine leukemia virus gp70 and bovine leukemia virus p24. Detection by radioimmunoassay of antibodies directed against these antigens.
Portetelle, Daniel ; ; et al
in Burny, Arsène (Ed.) Bovine leukosis: various methods of molecular virology (1977)Detailed reference viewed: 9 (1 ULg)
Purification of high molecular weight bacteriocin.
; ; Thonart, Philippe
Poster (1995, February)Detailed reference viewed: 5 (1 ULg)
Purification of mouse monoclonal antibodies from ascitic fluid by DEAE Affi-gel Blue chromatography.
; ; et al
in Langone, J.; Van Vunakis, H. (Eds.) Methods in Enzymology-Immunochemical Techniques - Part J (1986)Detailed reference viewed: 22 (9 ULg)
Purification of myeloperoxidase from equine polymorphonuclear leucocytes.
Mathy, Marianne ; ; Grulke, Sigrid et al
in Canadian Journal of Veterinary Research = Revue Canadienne de Recherche Vétérinaire (1998), 62(2), 127-32
Increases of plasma concentrations of neutrophil myeloperoxidase (MPO) can be used as markers of polymorphonuclear leucocytes (PMN) activation in pathological situations (sepsis, acute lung injury, acute ... [more ▼]
Increases of plasma concentrations of neutrophil myeloperoxidase (MPO) can be used as markers of polymorphonuclear leucocytes (PMN) activation in pathological situations (sepsis, acute lung injury, acute inflammation). To develop an assay for measurement of plasma MPO in horses during the above-mentioned infectious and inflammatory conditions, MPO was purified from equine PMN isolated from blood anticoagulated with citrate. PMN were extracted in a saline milieu (0.2 M Na acetate, 1 M NaCl, pH 4.7) to eliminate most of cellular proteins. Pellets were then extracted in the same buffer containing cationic detergent (1% cetyltrimethyl ammonium bromide). The supernatant was further purified by ion exchange chromatography (Hiload S Sepharose HP column 0.5 x 26 cm, equilibrated with 25 mM Na acetate, 0.2 M NaCl, pH 4.7) with a NaCl gradient (until 1 M). Most of the peroxidase activity of MPO (spectrophotometrically measured by the oxidation of orthodianisidine by hydrogen peroxide) was eluted at 0.65 M NaCl. MPO was further purified by gel filtration chromatography (Sephacryl S 200 column 2.6 x 42 cm with 25 mM Na acetate, 0.2 M NaCl, pH 4.7). MPO (specific activity: 74.3 U/mg) was obtained with a yield of 30% from the detergent extraction supernatant. Electrophoresis (non-reducing conditions) showed 3 bands identified, by comparison with human MPO, (i) the mature tetrameric enzyme (150 kDa) with 2 light and 2 heavy subunits, (ii) the precursor form (88 kDa) and (iii) a form of the heavy subunit without the prosthetic heme group (40 kDa). The mature enzyme and its precursor were glycosylated and possessed peroxidase activity. Equine MPO showed strong similarities with human and bovine MPO, with an absorption peak at 430 nm (Soret peak) characteristic of ferrimyeloperoxidase. Enzymatic activity was pH dependent (optimal value at pH 5.5). [less ▲]Detailed reference viewed: 43 (1 ULg)
Purification of pectin from apple pomace juice by using sodium caseinate and characterisation of their binding by isothermal titration calorimetry
; ; Paquot, Michel et al
in Food Hydrocolloids (2012), 29Detailed reference viewed: 27 (2 ULg)
Purification of pregnancy-associated glycoproteins from late-pregnancy Bubalus bubalis placentas and development of a radioimmunoassay for pregnancy diagnosis in water buffalo females.
; Melo de Sousa, Noelita ; et al
in BMC Veterinary Research (2013), 9
BACKGROUND: Pregnancy-associated glycoproteins (PAGs) were first described as placental antigens present in the blood serum of the mother soon after implantation. Here, we describe the purification of ... [more ▼]
BACKGROUND: Pregnancy-associated glycoproteins (PAGs) were first described as placental antigens present in the blood serum of the mother soon after implantation. Here, we describe the purification of several pregnancy-associated glycoproteins from water buffalo placenta (wbPAGs). A specific radioimmunoassay (RIA) was developed for early pregnancy diagnosis in buffalo species. RESULTS: Amino-terminal microsequencing of immunoreactive placental proteins allowed the identification of eleven wbPAGs sequences [Swiss-Prot accession numbers: P86369 to P86379]. Three polyclonal antisera (AS#858, AS#859 and AS#860) were raised in rabbits against distinct wbPAG fractions. A new RIA (RIA-860) was developed and used to distinguish between pregnant (n = 33) and non-pregnant (n = 26) water buffalo females. CONCLUSIONS: Our results confirmed the multiplicity of PAG expression in buffalo placenta. In addition, the RIA-860 system was shown to be sensitive, linear, reproducible, accurate and specific in measuring PAG concentrations in buffalo plasma samples from Day 37 of gestation onwards. [less ▲]Detailed reference viewed: 7 (1 ULg)
Purification of Progelatinases A and B by Continuous-Elution Electrophoresis
; ; et al
in Protein Expression & Purification (1995), 6(4), 417-22
Progelatinases A and B were purified from HT1080-conditioned culture medium using a continuous-elution electrophoresis. Initially cell culture medium was ammonium sulfate precipitated. The concentrated ... [more ▼]
Progelatinases A and B were purified from HT1080-conditioned culture medium using a continuous-elution electrophoresis. Initially cell culture medium was ammonium sulfate precipitated. The concentrated proteins were affinity chromatographed on gelatin-Sepharose column. The bound gelatinases were eluted with electrophoresis sample buffer and subjected to continuous-elution electrophoresis. In a single run, under the standardized working conditions, the obtained fractions contained four purified enzymes--progelatinase A (M(r) 72,000), its activated forms (M(r) 62,000 and M(r) 59,000), and progelatinase B (M(r) 92,000). Moreover, the continuous-elution electrophoresis allowed the enzymes separation from their respective inhibitors--TIMP-1 (M(r) 28,500) and TIMP-2 (M(r) 21,000). The purified progelatinases A and B demonstrated high specific activities (150-200 U/micrograms). [less ▲]Detailed reference viewed: 16 (0 ULg)
Purification of soybean lipoxygenase isoenzyme-1 and characterization of its inhibition by 13-hydroperoxides.
Fauconnier, Marie-Laure ;
in Grasas y Aceites (1996), 47(4), 242-246Detailed reference viewed: 21 (3 ULg)
Purification of the recombinant beta2 toxin (CPB2) from an enterotoxaemic bovine Clostridium perfringens strain and production of a specific immune serum
; Filée, Patrice ; Galleni, Moreno et al
in Protein Expression & Purification (2007), 55(1), 119-131
Overgrowth of Clostridium perfringens clones with production of one or more of its toxin(s) results in diverse digestive and systemic pathologies in human and animals, such as cattle enterotoxaemia. The ... [more ▼]
Overgrowth of Clostridium perfringens clones with production of one or more of its toxin(s) results in diverse digestive and systemic pathologies in human and animals, such as cattle enterotoxaemia. The so-called beta2 toxin (CPB2) is the most recently described major toxin produced by C perfringens. In this study, the cpb2 ORF (cpb2FM) from a cattle C perfringens-associated enterotoxaemia was cloned and sequenced. The cpb2FM and its deduced nucleotide sequence clearly corresponded to the epb2 allele considered as "consensus" and not to "atypical" allele, despite its "non-porcine" origin. Expression assays of the recombinant toxin CPB2FM were performed in Escherichia coli and Bacillus subtilis with the expression vector pBLTS72, and by genomic integration by double recombination in B. subtilis. Highest level of production was obtained with the expression vector in B. subtilis 168 strain. The recombinant CPB2FM protein was purified and a specific rabbit polyclonal antiserum was produced. Polyclonal antibodies could detect CPB2 production in supernatants of C. perfringens from enterotoxaemic cattle. (C) 2007 Elsevier Inc. All rights reserved. [less ▲]Detailed reference viewed: 25 (6 ULg)
A purification procedure of the tetrodotoxin binding component coming from the electroplaxes of E. electricus.
Dandrifosse, Guy ; Grandfils, Christian ; et al
in Changeux, J.P.; Maelicke, A; Neumann, E (Eds.) Molecular Basis of Nerve Activity (1984)Detailed reference viewed: 8 (0 ULg)
A purification procedure of the tetrodotoxin binding component contained in the electroplaxes of E. electricus
Dandrifosse, Guy ; Grandfils, Christian ; Bettendorff, Lucien et al
in Changeux, J.P.; Maelicke, A.; Neumann, E. (Eds.) Molecular Basis of Nerve Activity (1985)Detailed reference viewed: 17 (3 ULg)
Purification, Characterization, and Nucleotide Sequence of the Thermolabile Alpha-Amylase from the Antarctic Psychrotroph Alteromonas Haloplanctis A23
Feller, Georges ; ; Deroanne, Christophe et al
in Journal of Biological Chemistry (1992), 267(8), 5217-21
The alpha-amylase excreted by the antarctic bacterium Alteromonas haloplanctis was purified and the corresponding amy gene was cloned and sequenced. N- and C-terminal amino acid sequencing were used to ... [more ▼]
The alpha-amylase excreted by the antarctic bacterium Alteromonas haloplanctis was purified and the corresponding amy gene was cloned and sequenced. N- and C-terminal amino acid sequencing were used to establish the primary structure of the mature A. haloplanctis alpha-amylase which is composed of 453 amino acids with a predicted Mr of 49,340 and a pI of 5.5. Three Ca2+ ions are bound per molecule and its activity is modulated by chloride ions. Within the four consensus sequences, Asp-174, Glu-200, and Asp-264 are the proposed catalytic residues. The psychrotrophic A. haloplanctis alpha-amylase is characterized by a high amylolytic activity at low temperatures, a reduced apparent optimal temperature, and typical thermodynamic activation parameters A. haloplanctis alpha-amylase has also a low thermal stability as demonstrated by the temperature effect on both activity and secondary structure. It is suggested that structure flexibility and lower sensitivity of secondary structure to temperature variations in the low temperature range are the main structural adaptations of the psychrotrophic enzyme. The unusual stacking of small amino acids around the catalytic residues is proposed as a factor inducing active site flexibility and concomitant high activity of the enzyme at low temperatures. [less ▲]Detailed reference viewed: 17 (3 ULg)
Purification, expansion, and multiple fluorochrome labeling of cord blood hemopoietic precursors: preliminary results.
; Gothot, André ; Grosdent, Jean-Claude et al
in Journal of Hematotherapy (1993), 2(2), 259-61
CD34-positive cells were isolated from a total of 23 cords using CellPro Ceprate columns. AIS MicroCellector flasks, and panning. The cells were (1) expanded in serum-free culture supplemented with a ... [more ▼]
CD34-positive cells were isolated from a total of 23 cords using CellPro Ceprate columns. AIS MicroCellector flasks, and panning. The cells were (1) expanded in serum-free culture supplemented with a variety of combinations of cytokines and (2) immunophenotyped using multiple fluorochrome labeling. The results indicated that the avidin column produced the highest purity of CD34-positive cells, and that immature blast cells could be expanded in serum-free culture. Preliminary results suggested that the four fluorochrome labeling technique may provide useful information on the lineage commitment of cord blood precursor and blast cells. [less ▲]Detailed reference viewed: 24 (1 ULg)
Purification, refolding and characterization of the trimeric Omp2a outer membrane porin from Brucella melitensis.
; Matagne, André ; et al
in Protein Expression & Purification (2012), 83(2), 198-204
Brucella melitensis is a gram-negative bacteria known to cause brucellosis and to produce severe infections in humans. Whilst brucella's outer membrane proteins have been extensively studied due to their ... [more ▼]
Brucella melitensis is a gram-negative bacteria known to cause brucellosis and to produce severe infections in humans. Whilst brucella's outer membrane proteins have been extensively studied due to their potential role as antigens or virulence factors, their function is still poorly understood at the structural level, as the 3D structure of Brucella beta-barrel membrane proteins are still unknown. In this context, the B. melitensis trimeric Omp2a porin has been overexpressed and refolded in n-dodecyl-beta-d-maltopyranoside. We here show that this refolding process is insensitive to urea but is temperature- and ionic strength-dependent. Reassembled species were characterized by fluorescence, size-exclusion chromatography and circular dichroism. A refolding mechanism is proposed, suggesting that Omp2a first refolds under a monomeric form and then self-associates into a trimeric state. This first complete in vitro refolding of a membrane protein from B. melitensis shall eventually lead to functional and 3D structure determination. [less ▲]Detailed reference viewed: 19 (0 ULg)
Purificazione delle proteine associate alla gestazione (PAG) nella bufala (Bubalus bubalis): risultati preliminari.
; Melo de Sousa, Noelita ; et al
in Atti 2o Congresso Nazionale sull’Allevamento del Bufalo (2003)Detailed reference viewed: 13 (0 ULg)
Purified FSH supplemented with defined amounts of LH for superovulation in dairy cattle .
; Beckers, Jean-François ; et al
in Theriogenology (1988, January), 29(1), 260Detailed reference viewed: 12 (0 ULg)
Le purisme enfantin au prisme du discours spontané
in Français Moderne (Le) (2008), LXXXVI/1Detailed reference viewed: 19 (0 ULg)
Purpose and scope
; ; et al
in Nephrology Dialysis Transplantation (2013), 28Detailed reference viewed: 16 (2 ULg)
Purpura thrombotique thrombocytopénique
; ; et al
in Revue Médicale de Liège (1989), 44
Deux patientes âgées respectivement de 58 et 24 ans ont été hospitalisées dans le Service d'Hématologie du CHU Sart Tilman pour PTT. Le diagnostic a été établi à partir de certains symptômes classiques à ... [more ▼]
Deux patientes âgées respectivement de 58 et 24 ans ont été hospitalisées dans le Service d'Hématologie du CHU Sart Tilman pour PTT. Le diagnostic a été établi à partir de certains symptômes classiques à savoir : hémolyse intravasculaire avec schizocytose (40/1000 dans le premier cas et 30/1000 dans le deuxième), élévation importante des LDH, thrombocytopénie majeure < 30.000/mm³, signes de souffrance du système nerveux central et insuffisance rénale aiguë dans le premier cas. Le scanner abdominal a permis d'observer dans le premier cas (admis à un stade avancé) un infarctus splénique massif et dans les deux cas une augmentation des indices de résistivité artérielle, témoignant d'une souffrance rénale. Le traitement par échanges plasmatiques a été instauré avec succès. Après obtention de la rémission complète, des infusions de plasma frais ont été poursuivies et le traitement d'entretien a été relayé par les antiagrégants plaquettaires. [less ▲]Detailed reference viewed: 56 (2 ULg)