Humanization of yeast to produce complex terminally sialylated glycoproteins
Abstract
Yeast is a widely used recombinant protein expression system. We expanded its utility by engineering the yeast Pichia pastoris to secrete human glycoproteins with fully complex terminally sialylated N-glycans. After the knockout of four genes to eliminate yeast-specific glycosylation, we introduced 14 heterologous genes, allowing us to replicate the sequential steps of human glycosylation. The reported cell lines produce complex glycoproteins with greater than 90% terminal sialylation. Finally, to demonstrate the utility of these yeast strains, functional recombinant erythropoietin was produced.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
A Pichia pastoris yeast strain was engineered to generate human-like glycans but devoid of core-fucose.
“For example, a Pichia pastoris yeast strain was engineered to generate human-like glycans but devoid of core-fucose (Choi et al., 2003;Hamilton et al., 2006;Hamilton and Gerngross 2007).”
From Therapeutic glycoprotein production in mammalian cells (2017) · cites this paper as Hamilton et al., 2006
Cites
Carbohydrate-Specific Receptors of the Liver
Ashwell, G., & Harford, J. (1982) · Annual Review of Biochemistry · doi:10.1146/annurev.bi.51.070182.002531 · Cited by 1,914 · see citing works
Overview of N- and O-linked oligosaccharide structures found in various yeast species
Gemmill, T. R., & Trimble, R. B. (1999) · Biochimica et Biophysica Acta (BBA) - General Subjects · doi:10.1016/s0304-4165(98)00126-3 · Cited by 439 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Properties, production, and applications of camelid single-domain antibody fragments
Harmsen, M. M., & De Haard, H. J. (2007) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-007-1142-2 · Cited by 809 · see citing works
“However, this may now be feasible using P. pastoris strains with an engineered glycosylation machinery that are able to produce proteins with a specific human glycoform ( Hamilton et al. 2006 ).”
BackgroundTherapeutic glycoprotein production in mammalian cells
Lalonde, M.-E., & Durocher, Y. (2017) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2017.04.028 · Cited by 296 · see citing works
Engineered CHO cells for production of diverse, homogeneous glycoproteins
Yang (2015) · Nature Biotechnology · doi:10.1038/nbt.3280 · Cited by 254 · see citing works
Glycodelete engineering of mammalian cells simplifies N-glycosylation of recombinant proteins
Meuris, L., Santens, F., Elson, G., Festjens, N., Boone, M., Dos Santos, A., Devos, S., Rousseau, F., Plets, E., Houthuys, E., et al. (2014) · Nature Biotechnology · doi:10.1038/nbt.2885 · Cited by 141 · see citing works
Corchero, J. L., Gasser, B., Resina, D., Smith, W., Parrilli, E., Vázquez, F., Abasolo, I., Giuliani, M., Jäntti, J., Ferrer, P., et al. (2013) · Biotechnology Advances · doi:10.1016/j.biotechadv.2012.09.001 · Cited by 133 · see citing works
Engineering of complex protein sialylation in plants
Kallolimath, S., Castilho, A., Strasser, R., Grünwald-Gruber, C., Altmann, F., Strubl, S., Galuska, C. E., Zlatina, K., Galuska, S. P., Werner, S., et al. (2016) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1604371113 · Cited by 113 · see citing works
Amann, T., Schmieder, V., Faustrup Kildegaard, H., Borth, N., & Andersen, M. R. (2019) · Biotechnology and Bioengineering · doi:10.1002/bit.27101 · Cited by 46 · see citing works
- Authors
- Hamilton, S. R., Davidson, R. C., Sethuraman, N., Nett, J. H., Jiang, Y., Rios, S., Bobrowicz, P., Stadheim, T. A., Li, H., Choi, B.-K., Hopkins, D., Wischnewski, H., Roser, J., Mitchell, T., Strawbridge, R. R., Hoopes, J., Wildt, S., & Gerngross, T. U.
- Affiliations
- Dartmouth College (US)
- Corresponding author
- Gerngross, Tillman U.
- Venue
- Science
- Published
- 2006-09-08
- Volume
- 313
- Issue
- 5792
- Pages
- 1441-1443
- ISSN
- 0036-8075 (ISSN-L)
- 1095-9203
- OpenAlex
- W2107632694
- Cited by
- 511
- References
- 2
- Citations by year
- 2026: 18 · 2025: 10 · 2024: 8 · 2023: 9 · 2022: 11 · 2021: 14 · 2020: 25 · 2019: 17 · 2018: 21 · 2017: 30 · 2016: 28 · 2015: 28 · 2014: 33 · 2013: 34 · 2012: 42
- FWCI
- 15.21
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Science (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glycosylation and Glycoproteins Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
- Keywords
- Pichia pastoris, complex N-glycans, recombinant protein expression, erythropoietin, gene knockout, glycoprotein secretion
- MeSH terms
- Animals, Cell Line, Cloning, Molecular, Cytidine Monophosphate N-Acetylneuraminic Acid, Erythropoietin, Genetic Vectors, Glycosylation, Humans, Pichia, Recombinant Proteins, Sialic Acids, Sialoglycoproteins, Transformation, Genetic, Protein Engineering, Rats
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
