Apolipoprotein E Expression and Purification
Abstract
Since the discovery of the association of apolipoprotein E (apoE) 4 with Alzheimer's disease 17 years ago, numerous in vitro experiments with the apoE isoforms (apoE2, apoE3, and apoE4) have been performed to try to understand the basis for this association. The majority of these studies used commercial sources for apoE, but some used recombinant protein. In either case, these studies were most often conducted without considering the ramifications of the structural and biophysical differences among the three isoforms or without adequate quality control of the preparations. Here, we present a protocol for producing recombinant apoE that we have used successfully in our laboratory for the last 20 years. We also review the considerations that are critical for obtaining reliable and interpretable results with the end product.
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Bacterial ApoE is expressed as inclusion bodies that must be denatured, refolded, and optionally converted into LNPs using non-physiological procedures.
“Bacterial ApoE is expressed as inclusion bodies that must be denatured, refolded, and optionally converted into LNPs using non-physiological procedures, resulting in holoprotein that is different from its brain-derived counterpart with respect to key physical (e.g., lipid composition) and/or chemical (e.g., post-translational modifications) properties.21”
From ApoE is Secreted as a Lipid Nanoparticle by Mammalian Cells: Implications for Alzheimer’s Disease Pathogenesis (2025) · cites this paper as [21]
Bacterial ApoE holoprotein is different from its brain-derived counterpart with respect to key physical and/or chemical properties.
“resulting in holoprotein that is different from its brain-derived counterpart with respect to key physical (e.g., lipid composition) and/or chemical (e.g., post-translational modifications) properties.21”
From ApoE is Secreted as a Lipid Nanoparticle by Mammalian Cells: Implications for Alzheimer’s Disease Pathogenesis (2025) · cites this paper as [21]
To date all reports describing the biochemical properties of this recombinant protein have deployed lipid-free biosynthetic materials.
“Although ApoE is not detected in appreciable quantities in its apo form in physiological fluids, to date all reports describing the biochemical properties of this recombinant protein have deployed lipid-free biosynthetic materials.21–25”
From ApoE is Secreted as a Lipid Nanoparticle by Mammalian Cells: Implications for Alzheimer’s Disease Pathogenesis (2025) · cites this paper as [21]
ApoE is not detected in appreciable quantities in its apo form in physiological fluids.
“Although ApoE is not detected in appreciable quantities in its apo form in physiological fluids, to date all reports describing the biochemical properties of this recombinant protein have deployed lipid-free biosynthetic materials.21–25”
From ApoE is Secreted as a Lipid Nanoparticle by Mammalian Cells: Implications for Alzheimer’s Disease Pathogenesis (2025) · cites this paper as [21]
Cites
Apolipoprotein E structure: insights into function
Hatters, D. M., Peters-Libeu, C. A., & Weisgraber, K. H. (2006) · Trends in Biochemical Sciences · doi:10.1016/j.tibs.2006.06.008 · Cited by 583 · see citing works
Self-citationApolipoprotein E4 Forms a Molten Globule
Morrow, J. A., Hatters, D. M., Lu, B., Höchtl, P., Oberg, K. A., Rupp, B., & Weisgraber, K. H. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m204898200 · Cited by 204 · see citing works
Self-citationAmino-terminal domain stability mediates apolipoprotein E aggregation into neurotoxic fibrils
Hatters, D. M., Zhong, N., Rutenber, E., & Weisgraber, K. H. (2006) · Journal of Molecular Biology · doi:10.1016/j.jmb.2006.06.080 · Cited by 105 · see citing works
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Hernandez Arriaza, R., Reil, D., Fatuzzo, N., Fu, M., Dai, Y., Fernandez Martinez, D., Jiang, H., Holtzman, D. M., Greicius, M. D., & Khosla, C. (2025) · Biochemistry · doi:10.1021/acs.biochem.5c00503 · Cited by 2 · see citing works
- Authors
- Newhouse, Y., & Weisgraber, K. H.
- Affiliations
- University of California, San Francisco (US)
- Venue
- Methods in Molecular Biology
- Publisher
- Humana Press
- Published
- 2010
- Volume
- 670
- Pages
- 127-140
- ISSN
- 1064-3745 (ISSN-L)
- 1940-6029
- OpenAlex
- W2404122114
- Cited by
- 8
- References
- 3
- Citations by year
- 2026: 1 · 2025: 1 · 2023: 1 · 2022: 1 · 2014: 2 · 2012: 1
- FWCI
- 2.15
- Type
- book chapter
- Language
- en
- Open access
- closed
- Locations
- Methods in molecular biology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institute on Aging (R01AG028793)
- Topics
- Health Sciences › Medicine › Physiology › Alzheimer's disease research and treatments
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
- Health Sciences › Medicine › Surgery › Lipoproteins and Cardiovascular Health
- Keywords
- apolipoprotein E, recombinant protein expression, protein purification, APOE isoforms, APOE ε4, Alzheimer's disease, protein quality control
- MeSH terms
- Alzheimer Disease, Apolipoproteins E, Bacteria, Chromatography, Gel, Electrophoresis, Polyacrylamide Gel, Recombinant Proteins, Apolipoprotein E3, Apolipoprotein E4, Apolipoprotein E2
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