Document Detail


Reversed-phase high-performance liquid chromatography method for the determination of prolactin in bacterial extracts and in its purified form.
MedLine Citation:
PMID:  12075926     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Carlos R J Soares; Iara M C Camargo; Ligia Morganti; Elizabeth Gimbo; João Ezequiel de Oliveira; Richard Legoux; Pascual Ferrara; Paolo Bartolini
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of chromatography. A     Volume:  955     ISSN:  0021-9673     ISO Abbreviation:  J Chromatogr A     Publication Date:  2002 May 
Date Detail:
Created Date:  2002-06-21     Completed Date:  2002-12-12     Revised Date:  2009-01-15    
Medline Journal Info:
Nlm Unique ID:  9318488     Medline TA:  J Chromatogr A     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  229-36     Citation Subset:  IM    
Affiliation:
Biotechnology Department, IPEN-CNEN, Cidade Universitária, São Paulo, Brazil.
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MeSH Terms
Descriptor/Qualifier:
Bacteria / chemistry*
Blotting, Western
Chromatography, High Pressure Liquid / methods*
Electrophoresis, Polyacrylamide Gel
Humans
Prolactin / analysis*,  isolation & purification
Recombinant Proteins / analysis,  isolation & purification
Reproducibility of Results
Chemical
Reg. No./Substance:
0/Recombinant Proteins; 9002-62-4/Prolactin

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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