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Cited by 17Viral Infectious Diseases and Gene Expression in InsectsGlycosylation and Glycoproteins ResearchMicrobial Metabolic Engineering and Bioproduction

Prediction of N-linked glycoform profiles of monoclonal antibody with extracellular metabolites and two-step intracellular models

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Sha, Sha, Huang, Zhuangrong, Agarabi, Cyrus, Lute, Scott, Brorson, Kurt, Yoon, Seongkyu·2019·Processes
DOI 10.3390/pr7040227
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Abstract

Monoclonal antibodies (mAbs) are commonly glycosylated and show varying levels of galactose attachment. A set of experiments in our work showed that the galactosylation level of mAbs was altered by the culture conditions of hybridoma cells. The uridine diphosphate galactose (UDP-Gal) is one of the substrates of galactosylation. Based on that, we proposed a two-step model to predict N-linked glycoform profiles by solely using extracellular metabolites from cell culture. At the first step, the flux level of UDP-Gal in each culture was estimated based on a computational flux balance analysis (FBA); its level was found to be linear with the galactosylation degree on mAbs. At the second step, the glycoform profiles especially for G0F (agalactosylated), G1F (monogalactosylated) and G2F (digalactosylated) were predicted by a kinetic model. The model outputs well matched with the experimental data. Our study demonstrated that the integrated mathematical approach combining FBA and kinetic model is a promising strategy to predict glycoform profiles for mAbs during cell culture processes.

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  • Two-step frameworks for indirect NSD inference rely on pseudo-steady-state assumptions and therefore do not explicitly capture the transient dynamics of intracellular NSD pools over the course of a fed-batch culture.

    “Although this approach enables indirect inference of NSDs from extracellular data and alleviates some parametrisation challenges, it relies on pseudo-steadystate assumptions.It therefore does not explicitly capture the transient dynamics of intracellular NSD pools over the course of a fed-batch culture.”

    From Soft sensing of intracellular states for CHO cell bioprocessing with Ensemble Kalman Filters (2026) · cites this paper as [27]

    The Introduction and §3.2 describe a two-step model that infers nucleotide-sugar availability from extracellular data using FBA and adjusts Golgi concentrations between cultures, rather than tracking intracellular pool concentrations over time. The Discussion says this reduces kinetic parameters, but our copy does not explicitly state a pseudo-steady-state assumption. It also says the model was applied only to the exponential phase and proposes extending it to the stationary phase common in fed-batch cultures, so the fed-batch scope is not established. (LLM, conf=0.82)

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Cited by

Sorted by citations

Authors
Sha, S., Huang, Z., Agarabi, C., Lute, S., Brorson, K., & Yoon, S.
Affiliations
  • University of Massachusetts Lowell (US)
  • United States Food and Drug Administration (US)
Corresponding author
Yoon, Seongkyu
ORCID
Venue
Processes
Publisher
MDPI AG
Published
2019-04-23
Volume
7
Issue
4
Pages
227
ISSN
2227-9717 (ISSN-L)
DOI
10.3390/pr7040227
OpenAlex
W2941748144
Cited by
17
References
2
Citations by year
2026: 1 · 2025: 3 · 2023: 2 · 2022: 2 · 2021: 5 · 2020: 4
FWCI
0.90
Type
journal article
Language
en
Open access
Licence
Locations
  • Processes (publishedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
National Science Foundation (1706731)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glycosylation and Glycoproteins Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
Keywords
monoclonal antibodies, N-glycosylation, galactosylation, UDP-galactose, extracellular metabolites, flux balance analysis, kinetic model, hybridoma cell culture

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