Isolation of bovine plasma albumin by liquid chromatography and its polymerization for use in immunohematology

Detalhes bibliográficos
Autor(a) principal: Tanaka,K.
Data de Publicação: 2001
Outros Autores: Sawatani,E., Shigueoka,E.M., Dias,G.A., Nakao,H.C., Arashiro,F.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Brazilian Journal of Medical and Biological Research
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2001000800002
Resumo: The aim of the method described here is to remove hemoglobin, the major contaminant in the bovine plasma obtained from slaughterhouses, by adding a mixture of 19% cold ethanol and 0.6% chloroform, followed by fibrinogen and globulin precipitation by the Cohn method and nonspecific hemagglutinin by thermocoagulation. The experimental volume of bovine plasma was 2,000 ml per batch. Final purification was performed by liquid chromatography using the ion-exchange gel DEAE-Sepharose FF. The bovine albumin thus obtained presented > or = 99% purity, a yield of 25.0 ± 1.2 g/l plasma and >71.5% recovery. N-acetyl-DL-tryptophan (0.04 mmol/g protein) and sodium caprylate (0.04 mmol/g protein) were used as stabilizers and the final concentration of albumin was adjusted to 22.0% (w/v), pH 7.2 to 7.3. Viral inactivation was performed by pasteurization for 10 h at 60°C. The bovine albumin for the hemagglutination tests used in immunohematology was submitted to chemical treatment with 0.06% (w/v) glutaraldehyde and 0.1% (w/v) formaldehyde at 37°C for 12 h to obtain polymerization. A change in molecular distribution was observed after this treatment, with average contents of 56.0% monomers, 23.6% dimers, 12.2% trimers and 8.2% polymers. The tests performed demonstrated that this polymerized albumin enhances the agglutination of Rho(D)-positive red cells by anti-Rho(D) serum, permitting and improving visualization of the results.
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spelling Isolation of bovine plasma albumin by liquid chromatography and its polymerization for use in immunohematologybovine albumin, productionbovine albumin, purificationbovine albumin, polymerizationbovine albumin, downstream processThe aim of the method described here is to remove hemoglobin, the major contaminant in the bovine plasma obtained from slaughterhouses, by adding a mixture of 19% cold ethanol and 0.6% chloroform, followed by fibrinogen and globulin precipitation by the Cohn method and nonspecific hemagglutinin by thermocoagulation. The experimental volume of bovine plasma was 2,000 ml per batch. Final purification was performed by liquid chromatography using the ion-exchange gel DEAE-Sepharose FF. The bovine albumin thus obtained presented > or = 99% purity, a yield of 25.0 ± 1.2 g/l plasma and >71.5% recovery. N-acetyl-DL-tryptophan (0.04 mmol/g protein) and sodium caprylate (0.04 mmol/g protein) were used as stabilizers and the final concentration of albumin was adjusted to 22.0% (w/v), pH 7.2 to 7.3. Viral inactivation was performed by pasteurization for 10 h at 60°C. The bovine albumin for the hemagglutination tests used in immunohematology was submitted to chemical treatment with 0.06% (w/v) glutaraldehyde and 0.1% (w/v) formaldehyde at 37°C for 12 h to obtain polymerization. A change in molecular distribution was observed after this treatment, with average contents of 56.0% monomers, 23.6% dimers, 12.2% trimers and 8.2% polymers. The tests performed demonstrated that this polymerized albumin enhances the agglutination of Rho(D)-positive red cells by anti-Rho(D) serum, permitting and improving visualization of the results.Associação Brasileira de Divulgação Científica2001-08-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2001000800002Brazilian Journal of Medical and Biological Research v.34 n.8 2001reponame:Brazilian Journal of Medical and Biological Researchinstname:Associação Brasileira de Divulgação Científica (ABDC)instacron:ABDC10.1590/S0100-879X2001000800002info:eu-repo/semantics/openAccessTanaka,K.Sawatani,E.Shigueoka,E.M.Dias,G.A.Nakao,H.C.Arashiro,F.eng2001-07-20T00:00:00Zoai:scielo:S0100-879X2001000800002Revistahttps://www.bjournal.org/https://old.scielo.br/oai/scielo-oai.phpbjournal@terra.com.br||bjournal@terra.com.br1414-431X0100-879Xopendoar:2001-07-20T00:00Brazilian Journal of Medical and Biological Research - Associação Brasileira de Divulgação Científica (ABDC)false
dc.title.none.fl_str_mv Isolation of bovine plasma albumin by liquid chromatography and its polymerization for use in immunohematology
title Isolation of bovine plasma albumin by liquid chromatography and its polymerization for use in immunohematology
spellingShingle Isolation of bovine plasma albumin by liquid chromatography and its polymerization for use in immunohematology
Tanaka,K.
bovine albumin, production
bovine albumin, purification
bovine albumin, polymerization
bovine albumin, downstream process
title_short Isolation of bovine plasma albumin by liquid chromatography and its polymerization for use in immunohematology
title_full Isolation of bovine plasma albumin by liquid chromatography and its polymerization for use in immunohematology
title_fullStr Isolation of bovine plasma albumin by liquid chromatography and its polymerization for use in immunohematology
title_full_unstemmed Isolation of bovine plasma albumin by liquid chromatography and its polymerization for use in immunohematology
title_sort Isolation of bovine plasma albumin by liquid chromatography and its polymerization for use in immunohematology
author Tanaka,K.
author_facet Tanaka,K.
Sawatani,E.
Shigueoka,E.M.
Dias,G.A.
Nakao,H.C.
Arashiro,F.
author_role author
author2 Sawatani,E.
Shigueoka,E.M.
Dias,G.A.
Nakao,H.C.
Arashiro,F.
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Tanaka,K.
Sawatani,E.
Shigueoka,E.M.
Dias,G.A.
Nakao,H.C.
Arashiro,F.
dc.subject.por.fl_str_mv bovine albumin, production
bovine albumin, purification
bovine albumin, polymerization
bovine albumin, downstream process
topic bovine albumin, production
bovine albumin, purification
bovine albumin, polymerization
bovine albumin, downstream process
description The aim of the method described here is to remove hemoglobin, the major contaminant in the bovine plasma obtained from slaughterhouses, by adding a mixture of 19% cold ethanol and 0.6% chloroform, followed by fibrinogen and globulin precipitation by the Cohn method and nonspecific hemagglutinin by thermocoagulation. The experimental volume of bovine plasma was 2,000 ml per batch. Final purification was performed by liquid chromatography using the ion-exchange gel DEAE-Sepharose FF. The bovine albumin thus obtained presented > or = 99% purity, a yield of 25.0 ± 1.2 g/l plasma and >71.5% recovery. N-acetyl-DL-tryptophan (0.04 mmol/g protein) and sodium caprylate (0.04 mmol/g protein) were used as stabilizers and the final concentration of albumin was adjusted to 22.0% (w/v), pH 7.2 to 7.3. Viral inactivation was performed by pasteurization for 10 h at 60°C. The bovine albumin for the hemagglutination tests used in immunohematology was submitted to chemical treatment with 0.06% (w/v) glutaraldehyde and 0.1% (w/v) formaldehyde at 37°C for 12 h to obtain polymerization. A change in molecular distribution was observed after this treatment, with average contents of 56.0% monomers, 23.6% dimers, 12.2% trimers and 8.2% polymers. The tests performed demonstrated that this polymerized albumin enhances the agglutination of Rho(D)-positive red cells by anti-Rho(D) serum, permitting and improving visualization of the results.
publishDate 2001
dc.date.none.fl_str_mv 2001-08-01
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2001000800002
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2001000800002
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/S0100-879X2001000800002
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv text/html
dc.publisher.none.fl_str_mv Associação Brasileira de Divulgação Científica
publisher.none.fl_str_mv Associação Brasileira de Divulgação Científica
dc.source.none.fl_str_mv Brazilian Journal of Medical and Biological Research v.34 n.8 2001
reponame:Brazilian Journal of Medical and Biological Research
instname:Associação Brasileira de Divulgação Científica (ABDC)
instacron:ABDC
instname_str Associação Brasileira de Divulgação Científica (ABDC)
instacron_str ABDC
institution ABDC
reponame_str Brazilian Journal of Medical and Biological Research
collection Brazilian Journal of Medical and Biological Research
repository.name.fl_str_mv Brazilian Journal of Medical and Biological Research - Associação Brasileira de Divulgação Científica (ABDC)
repository.mail.fl_str_mv bjournal@terra.com.br||bjournal@terra.com.br
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