Title : O-linked glycosylation analysis of recombinant human
granulocyte colony-stimulating factor produced in glycoengineered Pichia pastoris by liquid chromatography and mass spectrometry
Abstract :
- Glycosylation is a major biochemical attribute of therapeutic proteins and detailed analyses including the structures and sites of such modifications are often required for product quality control and assurance
- Using liquid chromatography and tandem mass spectrometry techniques, we analyzed the O-linked glycosylation of recombinant human granulocyte colony-stimulating factor ( rhG- CSF ) derived from glycoengineered Pichia pastoris with regard to its nature, structure, occupancy, and location
- Peptide mappings using protease and chemical cleavages were performed to determine the specific O-linked glycosylation site used by Pichia-derived rhG- CSF
- Our results demonstrated that Thr134 , the equivalent O-linked glycosylation site found on endogenous human G-CSF , is the only site modified with a single mannose, allowing glycoengineered P. pastoris to be used as a viable production platform for therapeutic rhG- CSF