Rapid Characterization of Human Serum Albumin Binding for Per- and Polyfluoroalkyl Substances Using Differential Scanning Fluorimetry
Open accessPublished version of a preprint
Abstract
Abstract Per- and polyfluoroalkyl substances (PFAS) are a diverse class of synthetic chemicals that accumulate in the environment. Many proteins, including the primary human serum transport protein albumin (HSA), bind PFAS. The predictive power of physiologically based pharmacokinetic modeling approaches are currently limited by a lack of experimental data defining albumin binding properties for most PFAS. A novel thermal denaturation assay was optimized to evaluate changes in thermal stability of HSA in the presence of increasing concentrations of known ligands and a structurally diverse set of PFAS. Assay performance was initially evaluated for fatty acids and HSA binding drugs ibuprofen and warfarin. Concentration response relationships were determined and dissociation constants (K d ) for each compound were calculated using regression analysis of the dose-dependent changes in HSA melting temperature. Estimated K d values for HSA binding of octanoic acid, decanoic acid, hexadecenoic acid, ibuprofen and warfarin agreed with established values. The binding affinities for 24 PFAS that included perfluoroalkyl carboxylic acids (C4-C12), perfluoroalkyl sulfonic acids (C4-C8), mono- and polyether perfluoroalkyl ether acids, and polyfluoroalkyl fluorotelomer substances were determined. These results demonstrate the utility of this differential scanning fluorimetry assay as a rapid high through-put approach for determining the relative protein binding properties and identification of chemical structures involved in binding for large numbers of structurally diverse PFAS.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
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?
PFCAs with carbon chain lengths between 10 and 15 may bind to serum proteins and cross the blood-brain barrier in a similar way to saturated fatty acids.
“hypothesizing that PFCAs with carbon chain lengths between 10 and 15 may be binding to serum proteins and crossing the BBB in a similar way to saturated fatty acids.”
From A Critical Review and Meta-Analysis of Impacts of Per- and Polyfluorinated Substances on the Brain and Behavior (2022) · cites this paper as Jackson et al., 2021
The Discussion (page unavailable) says PFCAs resemble fatty acids and likely interact with fatty-acid binding sites on human serum albumin; the Results report albumin-binding measurements for PFCAs through C12. The paper does not establish binding across the claimed C10–C15 range or address blood–brain barrier crossing. It also reports that albumin binding is optimal at shorter chain lengths, rather than showing that C10–C15 PFCAs behave like saturated fatty acids throughout. (LLM, conf=0.88)
Experimental — accuracy not yet measuredIn vitro work demonstrates that proteins like human serum albumin and human L-FABP have optimal affinity for PFAS with carbon chain length around 10.
“This hypothesis is supported by in vitro work demonstrating that proteins like human serum albumin and human L-FABP have optimal affinity for PFAS with carbon chain length around 10 (Zhang et al., 2013; Jackson et al., 2021).”
From A Critical Review and Meta-Analysis of Impacts of Per- and Polyfluorinated Substances on the Brain and Behavior (2022) · cites this paper as Jackson et al., 2021
The Results report highest HSA affinity at 7–9 aliphatic carbons, and the Discussion says C10+ PFAS bind less well. Thus Jackson et al. support a chain-length optimum for HSA, but not an optimum at carbon length 10. The study examined HSA, not human L-FABP; the sentence also cites Zhang et al. for that combined claim. (LLM, conf=0.88)
Experimental — accuracy not yet measured
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Smoothing and Differentiation of Data by Simplified Least Squares Procedures.
Savitzky, A., & Golay, M. J. E. (1964) · Analytical Chemistry · doi:10.1021/ac60214a047 · Cited by 21,904 · see citing works
Buck, R. C., Franklin, J., Berger, U., Conder, J. M., Cousins, I. T., de Voogt, P., Jensen, A. A., Kannan, K., Mabury, S. A., & van Leeuwen, S. P. (2011) · Integrated Environmental Assessment and Management · doi:10.1002/ieam.258 · Cited by 4,641 · see citing works
Sources, fate and transport of perfluorocarboxylates
Prevedouros, K., Cousins, I. T., Buck, R. C., & Korzeniowski, S. H. (2006) · Environmental Science & Technology · doi:10.1021/es0512475 · Cited by 2,594 · see citing works
Niesen, F., Berglund, H., & Vedadi, M. (2007) · Nature Protocols · doi:10.1038/nprot.2007.321 · Cited by 2,420 · see citing works
Fatty acid binding to plasma albumin
Spector, A. A. (1975) · Journal of Lipid Research · doi:10.1016/s0022-2275(20)36723-7 · Cited by 970 · see citing works
Monitoring of Perfluorinated Compounds in Aquatic Biota: An Updated Review: PFCs in Aquatic Biota
Houde, M., De Silva, A. O., Muir, D. C. G., & Letcher, R. J. (2011) · Environmental Science & Technology · doi:10.1021/es104326w · Cited by 847 · see citing works
Thermofluor-based high-throughput stability optimization of proteins for structural studies
Ericsson, U. B., Hällberg, B. M., DeTitta, G. T., Dekker, N., & Nordlund, P. (2006) · Analytical Biochemistry · doi:10.1016/j.ab.2006.07.027 · Cited by 840 · see citing works
PFAS concentrations in soils: Background levels versus contaminated sites
Brusseau, M. L., Anderson, R. H., & Guo, B. (2020) · Science of The Total Environment · doi:10.1016/j.scitotenv.2020.140017 · Cited by 817 · see citing works
Evaluation of fluorescence-based thermal shift assays for hit identification in drug discovery
Lo, M.-C., AULABAUGH, A. E., Jin, G., Cowling, R. A., Bard, J., Malamas, M. S., & Ellestad, G. A. (2004) · Analytical Biochemistry · doi:10.1016/j.ab.2004.04.031 · Cited by 688 · see citing works
Vedadi, M., Niesen, F., Allali‐Hassani, A., Fedorov, O., Finerty, P. J., Wasney, G. A., Yeung, R., Arrowsmith, C. H., Ball, L., Berglund, H., et al. (2006) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0605224103 · Cited by 590 · see citing works
Matulis, D., Kranz, J. K., Salemme, F. R., & Todd, M. J. (2005) · Biochemistry · doi:10.1021/bi048135v · Cited by 495 · see citing works
Molecular origins of fluorocarbon hydrophobicity
Dalvi, V. H., & Rossky, P. J. (2010) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0915169107 · Cited by 429 · see citing works
Binding of Perfluorooctanoic Acid to Rat and Human Plasma Proteins
Han, X., Snow, T. A., Kemper, R. A., & Jepson, G. W. (2003) · Chemical Research in Toxicology · doi:10.1021/tx034005w · Cited by 411 · see citing works
Albumin is the major carrier protein for PFOS, PFOA, PFHxS, PFNA and PFDA in human plasma
Forsthuber, M., Kaiser, A. M., Granitzer, S., Hassl, I., Hengstschläger, M., Stangl, H., & Gundacker, C. (2020) · Environment International · doi:10.1016/j.envint.2019.105324 · Cited by 274 · see citing works
Theory and applications of differential scanning fluorimetry in early-stage drug discovery
Gao, K., Oerlemans, R., & Groves, M. R. (2020) · Biophysical Reviews · doi:10.1007/s12551-020-00619-2 · Cited by 269 · see citing works
Kotlarz, N., McCord, J., Collier, D., Lea, C. S., Strynar, M., Lindstrom, A. B., Wilkie, A. A., Islam, J. Y., Matney, K., Tarte, P., et al. (2020) · Environmental Health Perspectives · doi:10.1289/ehp6837 · Cited by 261 · see citing works
Self-citationWang, Z., Cousins, I. T., Scheringer, M., Buck, R. C., & Hungerbühler, K. (2014) · Environment International · doi:10.1016/j.envint.2014.04.006 · Cited by 242 · see citing works
Beesoon, S., & Martin, J. W. (2015) · Environmental Science & Technology · doi:10.1021/es505399w · Cited by 223 · see citing works
Noncovalent Interactions of Long-Chain Perfluoroalkyl Acids with Serum Albumin
Bischel, H. N., MacManus-Spencer, L. A., & Luthy, R. G. (2010) · Environmental Science & Technology · doi:10.1021/es101334s · Cited by 184 · see citing works
Fluorescence study on site-specific binding of perfluoroalkyl acids to human serum albumin
Chen, Y., & Guo, L. (2008) · Archives of Toxicology · doi:10.1007/s00204-008-0359-x · Cited by 183 · see citing works
Guillette, T. C., McCord, J., Guillette, M., Polera, M. E., Rachels, K. T., Morgeson, C., Kotlarz, N., Knappe, D. R. U., Reading, B. J., Strynar, M., et al. (2020) · Environment International · doi:10.1016/j.envint.2019.105358 · Cited by 164 · see citing works
Self-citationAdvances in molecular and cell biology
Bittar, E. E. (1992) · JAI Press eBooks · Cited by 162 · see citing works
Thermal Denaturation Assays in Chemical Biology
Senisterra, G. A., Chau, I., & Vedadi, M. (2011) · Assay and Drug Development Technologies · doi:10.1089/adt.2011.0390 · Cited by 148 · see citing works
Cheng, W., & Ng, C. A. (2018) · Environmental Science & Technology · doi:10.1021/acs.est.8b01268 · Cited by 141 · see citing works
Determination of Protein-ligand Interactions Using Differential Scanning Fluorimetry
Vivoli, M., Novak, H. R., Littlechild, J. A., & Harmer, N. J. (2014) · Journal of Visualized Experiments · doi:10.3791/51809 · Cited by 139 · see citing works
Determination of Energies and Sites of Binding of PFOA and PFOS to Human Serum Albumin
Salvalaglio, M., Muscionico, I., & Carlo, C. (2010) · The Journal of Physical Chemistry B · doi:10.1021/jp106584b · Cited by 128 · see citing works
Screening for Ligands Using a Generic and High-Throughput Light-Scattering-Based Assay
Senisterra, G. A., Markin, E., Yamazaki, K., Hui, R., Vedadi, M., & Awrey, D. E. (2006) · SLAS DISCOVERY · doi:10.1177/1087057106294699 · Cited by 126 · see citing works
Ng, C. A., & Hungerbühler, K. (2015) · Environmental Science & Technology · doi:10.1021/acs.est.5b03000 · Cited by 122 · see citing works
Ràfols, C., Amézqueta, S., Fuguet, E., et al. (2017) · Journal of Pharmaceutical and Biomedical Analysis · doi:10.1016/j.jpba.2017.12.008 · Cited by 101 · see citing works
Hebert, P. C., & MacManus‐Spencer, L. A. (2010) · Analytical Chemistry · doi:10.1021/ac100721e · Cited by 92 · see citing works
Simeonov, A. M. (2013) · Expert Opinion on Drug Discovery · doi:10.1517/17460441.2013.806479 · Cited by 91 · see citing works
Solvent Suitability for HFPO-DA (“GenX” Parent Acid) in Toxicological Studies
Liberatore, H. K., Jackson, S. R., Strynar, M. J., & McCord, J. P. (2020) · Environmental Science & Technology Letters · doi:10.1021/acs.estlett.0c00323 · Cited by 84 · see citing works
Cheng, W., & Ng, C. A. (2017) · Environmental Science & Technology · doi:10.1021/acs.est.7b02602 · Cited by 83 · see citing works
Interaction of perfluorooctanoic acid with human serum albumin
Wu, L.-L., Gao, H.-W., Gao, N.-Y., Chen, F.-F., & Chen, L. (2009) · BMC Structural Biology · doi:10.1186/1472-6807-9-31 · Cited by 81 · see citing works
Ràfols, C., Zarza, S., & Bosch, E. (2014) · Talanta · doi:10.1016/j.talanta.2014.06.060 · Cited by 78 · see citing works
Fluorotechnology Is Critical to Modern Life: The FluoroCouncil Counterpoint to the Madrid Statement
Bowman, J. S. (2015) · Environmental Health Perspectives · doi:10.1289/ehp.1509910 · Cited by 75 · see citing works
Quantitation of protein–protein interactions by thermal stability shift analysis
Layton, C. J., & Hellinga, H. W. (2011) · Protein Science · doi:10.1002/pro.674 · Cited by 73 · see citing works
Binding Properties of Hydrophobic Molecules to Human Serum Albumin Studied by Fluorescence Titration
Takehara, K., Yuki, K., Shirasawa, M., et al. (2009) · Analytical Sciences · doi:10.2116/analsci.25.115 · Cited by 39 · see citing works
The Ligand Affinity of Proteins Measured by Isothermal Denaturation Kinetics
Epps, D. E. V., Sarver, R. W., Rogers, J. M., et al. (2001) · Analytical Biochemistry · doi:10.1006/abio.2001.5047 · Cited by 37 · see citing works
Lee, I. Y., & McMenamy, R. H. (1980) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)43709-x · Cited by 34 · see citing works
Effects of Fluorinated and Hydrogenated Surfactants on Human Serum Albumin at Different pHs
Sabín, J., Prieto, G., González‐Pérez, A., Ruso, J. M., & Sarmiento, F. (2005) · Biomacromolecules · doi:10.1021/bm050549w · Cited by 32 · see citing works
Honoré, B., & Brodersen, R. (1988) · Analytical Biochemistry · doi:10.1016/0003-2697(88)90124-8 · Cited by 30 · see citing works
Weak Polar Interactions Confer Albumin Binding Site Selectivity for Haloether Anesthetics
Liu, R., & Eckenhoff, R. G. (2005) · Anesthesiology · doi:10.1097/00000542-200504000-00016 · Cited by 24 · see citing works
Vorum, H., Pedersen, A. O., & Honoré, B. (1992) · International journal of peptide & protein research · doi:10.1111/j.1399-3011.1992.tb00319.x · Cited by 12 · see citing works
Flagged first, then by citations
Research works printed but not linked: 1
- [6]Fourth meeting of the Conference of the Parties to the Stockholm Convention on Persistent Organic Pollutants. Establishing indicative elements of a work programme to facilitate the elimination of listed brominated diphenyl ethers and the restriction or elimination of perfluorooctane sulfonic acid and its salts, perfluorooctane sulfonyl fluoride and other chemicals listed in Annexes A or B of the Convention at the fourth meeting of the Conference of the Parties, 2009.
Cited by
Sorted by citations
Evans, N., Conley, J. M., Cardon, M., Hartig, P., Medlock-Kakaley, E., & Gray, L. E. (2022) · Toxicology and Applied Pharmacology · doi:10.1016/j.taap.2022.116136 · Cited by 208 · see citing works
Starnes, H. M., Rock, K. D., Jackson, T. W., & Belcher, S. M. (2022) · Frontiers in Toxicology · doi:10.3389/ftox.2022.881584 · Cited by 166 · see citing works
Self-citationPFAS Exposures and the Human Metabolome: A Systematic Review of Epidemiological Studies
India-Aldana, S., Yao, M., Midya, V., Colicino, E., Chatzi, L., Chu, J., Gennings, C., Jones, D. P., Loos, R. J. F., Setiawan, V. W., et al. (2023) · Current Pollution Reports · doi:10.1007/s40726-023-00269-4 · Cited by 114 · see citing works
Crisalli, A. M., Cai, A., & Cho, B. P. (2023) · Chemical Research in Toxicology · doi:10.1021/acs.chemrestox.3c00011 · Cited by 40 · see citing works
- Authors
- Jackson, T. W., Scheibly, C. M., Polera, M. E., & Belcher, S. M.
- Affiliations
- North Carolina State University (US)
- Corresponding author
- Belcher, Scott M.
- Venue
- Environmental Science & Technology
- Publisher
- American Chemical Society (ACS)
- Published
- 2021-09-08
- Volume
- 55
- Issue
- 18
- Pages
- 12291-12301
- ISSN
- 0013-936X (ISSN-L)
- 1520-5851
- PMC
- PMC8651256
- OpenAlex
- W3197518040
- Cited by
- 87
- References
- 44
- Citations by year
- 2026: 19 · 2025: 29 · 2024: 11 · 2023: 20 · 2022: 6 · 2021: 2
- FWCI
- 3.06
- Type
- journal article
- Language
- en
- Open access
- green
- doi.org
- Licence
- Locations
- Environmental Science & Technology (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- U.S. Department of Health and Human Services (P42ES031009, T32ES007046)
- North Carolina State University
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Interaction Studies and Fluorescence Analysis
- Physical Sciences › Environmental Science › Environmental Chemistry › Per- and polyfluoroalkyl substances research
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical Methods in Pharmaceuticals
- Keywords
- per- and polyfluoroalkyl substances, PFAS, human serum albumin, protein binding, differential scanning fluorimetry, thermal denaturation, dissociation constant, physiologically based pharmacokinetic modeling, perfluorinated carboxylic acids, perfluoroalkyl sulfonic acids, fatty acids, ibuprofen, warfarin, high-throughput binding assay
- MeSH terms
- Serum Albumin, Human, Carboxylic Acids, Fluorocarbons, Fluorometry, Humans, Sulfonic Acids, Alkanesulfonic Acids
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
