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A tandem mass spectrometric approach to the identification of O‐glycosylated glargine glycoforms in active pharmaceutical ingredient expressed in Pichia pastoris
Author(s) -
Kannan Vivekanandan,
Narayanaswamy Pradeep,
Gadamsetty Deepak,
Hazra Partha,
Khedkar Anand,
Iyer Harish
Publication year - 2009
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.3965
Subject(s) - chemistry , active ingredient , pichia pastoris , ingredient , chromatography , tandem mass spectrometry , identification (biology) , tandem , mass spectrometry , biochemistry , recombinant dna , pharmacology , food science , gene , medicine , botany , materials science , composite material , biology
Glycoforms of glargine expressed in Pichia pastoris were isolated by high‐performance liquid chromatography and analyzed by a series of chemical and mass spectrometric methods for the identification of various glycoforms, glycosylation position, nature and structure of glycans. Reduction and alkylation, peptide mapping techniques were used to decipher the amino acid site at which glycosylation had taken place. Chemical methods were coupled with mass spectrometry techniques such as electrospray ionization and matrix‐assisted laser desorption/ionization for identification of the glycosylation site. Copyright © 2009 John Wiley & Sons, Ltd.