Premium
N ‐glycans of recombinant human interferon‐γ change during batch culture of chinese hamster ovary cells
Author(s) -
Hooker Andrew D.,
Goldman Merlin H.,
Markham Nicola H.,
James David C.,
Ison Andrew P.,
Bull Alan T.,
Strange Philip G.,
Salmon Ian,
Baines Anthony J.,
Jenkins Nigel
Publication year - 1995
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260480612
Subject(s) - chinese hamster ovary cell , exoglycosidase , glycosylation , glycan , recombinant dna , cell culture , biochemistry , chemistry , biology , n linked glycosylation , microbiology and biotechnology , glycoprotein , genetics , gene
A recombinant Chinese hamster ovary (CHO) cell line making human interfron‐γ (IFN‐γ) was grown in 12‐L stirred tank fermentors in three batch fermentations under conditions of constant temperature, pH, and dissolved oxygen tension. In addition to cell growth, metabolite, and productivity data, a detailed analysis of the carbohydrate structures attached to each glycosylation site of IFN‐γ was achieved using matrix‐assisted laser desorption mass spectrometry (MALDI‐MS) in combination with exoglycosidase array sequencing. Complex biantennary oligosaccharides (particularly Gal 2 GlcNAc 4 Man 3 which was core ℵl‐6 fucosylated at Asn 25 but not at Asng 97 ) were most prevalent at both glycosylation sites. However, considerable microheterogeneity arising from the presence of triantennary and truncated glycan structures was also observed. The proportion of the dominant core glycan structure (Gal 2 GlcNAc 4 Man 3 ± Fuc 1 ) decreased by 15–26% during batch culture, with increases in the proportion of oligomannose and truncated glycans over the same time period. Prolonged culture resulting from an extended lag phase led to further accumulation of oligomannose and truncated structures, reaching up to 52% of total glycans attached to Asng 97 by 240 h of culture. The implications of these glycosylation changes for optimizing the time for harvesting cell cultures, and for the clearance of recombinant therapeutic products in vivo are discussed. © 1995 John Wiley & Sons, Inc.