TY - JOUR
T1 - A Markov chain model for N-linked protein glycosylation – towards a low-parameter tool for model-driven glycoengineering
AU - Spahn, Philipp N.
AU - Hansen, Anders Holmgaard
AU - Hansen, Henning Gram
AU - Arnsdorf, Johnny
AU - Kildegaard, Helene Faustrup
AU - Lewis, Nathan
PY - 2016
Y1 - 2016
N2 - Glycosylation is a critical quality attribute of most recombinant biotherapeutics. Consequently, drug development requires careful control of glycoforms to meet bioactivity and biosafety requirements. However, glycoengineering can be extraordinarily difficult given the complex reaction networks underlying glycosylation and the vast number of different glycans that can be synthesized in a host cell. Computational modeling offers an intriguing option to rationally guide glycoengineering, but the high parametric demands of current modeling approaches pose challenges to their application. Here we present a novel low-parameter approach to describe glycosylation using flux-balance and Markov chain modeling. The model recapitulates the biological complexity of glycosylation, but does not require user-provided kinetic information. We use this method to predict and experimentally validate glycoprofiles on EPO, IgG as well as the endogenous secretome following glycosyltransferase knock-out in different Chinese hamster ovary (CHO) cell lines. Our approach offers a flexible and user-friendly platform that can serve as a basis for powerful computational engineering efforts in mammalian cell factories for biopharmaceutical production.
AB - Glycosylation is a critical quality attribute of most recombinant biotherapeutics. Consequently, drug development requires careful control of glycoforms to meet bioactivity and biosafety requirements. However, glycoengineering can be extraordinarily difficult given the complex reaction networks underlying glycosylation and the vast number of different glycans that can be synthesized in a host cell. Computational modeling offers an intriguing option to rationally guide glycoengineering, but the high parametric demands of current modeling approaches pose challenges to their application. Here we present a novel low-parameter approach to describe glycosylation using flux-balance and Markov chain modeling. The model recapitulates the biological complexity of glycosylation, but does not require user-provided kinetic information. We use this method to predict and experimentally validate glycoprofiles on EPO, IgG as well as the endogenous secretome following glycosyltransferase knock-out in different Chinese hamster ovary (CHO) cell lines. Our approach offers a flexible and user-friendly platform that can serve as a basis for powerful computational engineering efforts in mammalian cell factories for biopharmaceutical production.
KW - Glycosylation
KW - Glycoengineering
KW - Markov chains
KW - Flux-balance analysis
U2 - 10.1016/j.ymben.2015.10.007
DO - 10.1016/j.ymben.2015.10.007
M3 - Journal article
C2 - 26537759
SN - 1096-7176
VL - 33
SP - 52
EP - 66
JO - Metabolic Engineering
JF - Metabolic Engineering
ER -