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Reversed-phase high-performance liquid chromatography method for the determination of prolactin in bacterial extracts and in its purified form.
Soares, Carlos R J; Camargo, Iara M C; Morganti, Ligia; Gimbo, Elizabeth; Ezequiel de Oliveira, João; Legoux, Richard; Ferrara, Pascual; Bartolini, Paolo.
Afiliação
  • Soares CR; Biotechnology Department, IPEN-CNEN, Cidade Universitária, São Paulo, Brazil.
J Chromatogr A ; 955(2): 229-36, 2002 May 10.
Article em En | MEDLINE | ID: mdl-12075926
Reversed-phase high-performance liquid chromatography methodology for the determination of human prolactin (hPRL) in bacterial periplasmic space or in purified preparations has been developed. The technique, based on the high hydrophobicity of the hPRL molecule, allows its separation from the bulk of bacterial proteins. The precision for periplasmic shock fluid analysis was characterized by relative standard variations of 3-7% for intra-day and of 3-25% for inter-day determinations. Accuracy, evaluated by recovery tests, was of the order of 90%, a calibration curve being constructed with the use of a lyophilized osmotic shock fluid extract, which provided a stable, readily prepared internal reference. Sensitivity was of the order of 0.5 microg of hPRL. The methodology developed also provided a tool for comparing the hydrophobicity of glycosylated and non-glycosylated prolactin molecules obtained from several different species and of different preparations of native or biosynthetic human prolactin.
Assuntos
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Prolactina / Bactérias / Cromatografia Líquida de Alta Pressão Limite: Humans Idioma: En Revista: J Chromatogr A Ano de publicação: 2002 Tipo de documento: Article País de afiliação: Brasil
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Prolactina / Bactérias / Cromatografia Líquida de Alta Pressão Limite: Humans Idioma: En Revista: J Chromatogr A Ano de publicação: 2002 Tipo de documento: Article País de afiliação: Brasil