Document Detail


The Separation of Recombinant Human Growth Hormone Variants by UHPLC.
MedLine Citation:
PMID:  23212133     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Reversed-phase ultra-high-performance liquid chromatography (RP-UHPLC) was used for the separation of recombinant human growth hormone (rhGH) variants. A bridged ethylene hybrid silica C18 column was used at 37°C. The composition and temperature of the mobile phase were optimized for the separation. An isocratic elution, with approximately 46% acetonitrile in 25 mM potassium borate buffer (pH 8.5), was found to give superior selectivity in comparison with commonly used mobile phases. The method separated eight rhGH variants: (i) di-oxy Met14/Met125 sulfoxide, (ii) Met125 sulfoxide, (iii) Met14 sulfoxide, (iv) mono-deamidated (Asn149 → Asp149 or Asn152 → Asp152), (v) di-deamidated (Asn149 → Asp149 and Asn152 → Asp152), (vi) clip (Thr142-Tyr143), (vii) desPhe1 and (viii) trisulfide (Cys182-SSS-Cys189) from each other and from the native rhGH. Characterization of the purified variants was conducted by liquid chromatography-mass spectrometry tryptic mapping. The novel mobile phase, in combination with the UHPLC system, generated a significantly higher resolution than previously reported reversed-phase LC methods, including pharmacopoeal methods, for analyzing rhGH.
Authors:
Göran Karlsson; Kathryn Eriksson; Annevi Persson; Håkan Månsson; Susanne Söderholm
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-12-4
Journal Detail:
Title:  Journal of chromatographic science     Volume:  -     ISSN:  1945-239X     ISO Abbreviation:  J Chromatogr Sci     Publication Date:  2012 Dec 
Date Detail:
Created Date:  2012-12-5     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0173225     Medline TA:  J Chromatogr Sci     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Pfizer Health AB, Box 108, SE-64541 Strängnäs, Sweden.
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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