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Author |
Rosa, P.A.J.; Azevedo, A.M.; Aires-Barros, M.R. |
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Title |
Application of central composite design to the optimisation of aqueous two-phase extraction of human antibodies |
Type |
Journal Article |
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Year |
2007 |
Publication |
Journal of Chromatography. A |
Abbreviated Journal |
J Chromatogr A |
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Volume |
1141 |
Issue |
1 |
Pages |
50-60 |
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Keywords |
Analysis of Variance; Animals; Antibodies/*chemistry/*isolation & purification; Buffers; Chemical Fractionation/*methods; Horses; Humans; Hydrophobicity; Isoelectric Point; Models, Biological; Molecular Weight; Myoglobin/chemistry/isolation & purification; Osmolar Concentration; Phase Transition; Polyethylene Glycols; Serum Albumin/chemistry/isolation & purification; Sodium Chloride |
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Abstract |
The partition of human antibodies in aqueous two-phase systems (ATPSs) of polyethylene glycol (PEG) and phosphate was systematically studied using first pure proteins systems and then an artificial mixture of proteins containing 1mg/ml human immunoglobulin G (IgG), 10mg/ml serum albumin and 2mg/ml myoglobin. Preliminary results obtained using pure proteins systems indicated that the PEG molecular weight and concentration, the pH value and the salts concentration had a pronounced effect on the partitioning behaviour of all proteins. For high ionic strengths and pH values higher than the isoelectric point (pI) of the contaminant proteins, IgG could be selectively recovered on the top phase. According to these results, a face centred composite design was performed in order to optimise the purification of IgG from the mixture of proteins. The optimal conditions for the isolation of IgG were observed for high concentrations of NaCl and low concentrations of both phase forming components. The best purification was achieved using an ATPS containing 8% (w/w) PEG 3350, 10% (w/w) phosphate pH 6 and 15% (w/w) NaCl. A recovery yield of 101+/-7%, a purity of 99+/-0% and a yield of native IgG of 97+/-4% were obtained. Back extraction studies of IgG to a new phosphate phase were performed and higher yields were obtained using 10% phosphate buffer at pH 6. The total extraction yield was 76% and the purity 100%. |
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Address |
IBB Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Instituto Superior Tecnico, Av Rovisco Pais, 1049-001 Lisbon, Portugal |
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English |
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ISSN |
0021-9673 |
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Notes |
PMID:17196214 |
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no |
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Call Number  |
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Serial |
1842 |
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Author |
Mejdell, C.M.; Simensen, E.; Boe, K.E. |
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Title |
Is snow a sufficient source of water for horses kept outdoors in winter? A case report |
Type |
Journal Article |
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Year |
2005 |
Publication |
Acta Veterinaria Scandinavica |
Abbreviated Journal |
Acta Vet Scand |
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Volume |
46 |
Issue |
1-2 |
Pages |
19-22 |
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Keywords |
Animals; *Drinking Behavior; *Horses; Norway; Osmolar Concentration; Snow; *Thirst; *Water Supply |
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Abstract |
Due to extreme weather conditions, a flock of outwintered Icelandic horses had to manage for several days on snow as the source of free water. They were fed grass silage ad lib, and any change in feed consumption was not observed. After nine days, blood samples were taken and analysed for plasma osmolality, they were subjected to a simple clinical examination, and offered drinking water. Osmolality levels were within normal limits and mean value did not differ significantly from samples which previously were taken of the same individuals. The general condition of the horses was normal, with no signs of clinical dehydration or disease. The horses showed very little interest for the offered drinking water. This suggests that in cold winter weather, horses being fed grass silage and adjusted to eat snow, can manage for several days with snow substituting liquid water without their physiology and welfare being challenged. |
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Address |
National Centre for Veterinary Contract Research and Commercial Services, Ltd |
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English |
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ISSN |
0044-605X |
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Notes |
PMID:16108209 |
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Call Number |
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Serial |
1886 |
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