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Dispersion of Nanoparticles in Different Media Importantly Determines the Composition of Their Protein Corona

Strojan, Klemen, Leonardi, Adrijana, Bregar, Vladimir B., Križaj, Igor, Svete, Jurij, Pavlin, Mojca·2017·PLOS ONE
DOI 10.1371/journal.pone.0169552PMC PMC5215476
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Fig 1 FTIR measurements. Samples were dried on the diamond crystal from solutions/dispersions prior to ATR-FTIR measurements. A: ATR-FTIR spectra of cobalt ferrite NPs (Co ferrite NPs), PAA, PAA NPs and silica NPs. B: ATR-FTIR spectra of cobalt ferrite NPs (Co ferrite NPs), PAA, and PAA NPs in the 1900–900 cm-1 region. C: ATR-FTIR spectrum of silica NPs in the 4000–600 cm-1 region.
Fig 1 FTIR measurements. Samples were dried on the diamond crystal from solutions/dispersions prior to ATR-FTIR measurements. A: ATR-FTIR spectra of cobalt ferrite NPs (Co ferrite NPs), PAA, PAA NPs and silica NPs. B: ATR-FTIR spectra of cobalt ferrite NPs (Co ferrite NPs), PAA, and PAA NPs in the 1900–900 cm-1 region. C: ATR-FTIR spectrum of silica NPs in the 4000–600 cm-1 region.CC BY · PubMed Central
Fig 2 Analysis of NPs protein corona using SDS-PAGE. PAA (A) and silica (B) NPs were dispersed in different media (dH2O, NaCl, PBS, RPMI) and incubated for 1 h in 10% or 100% FBS. Samples denoted with (+) contained PAA (A) or silica (B) NPs. In samples without NPs (-) were the proteins which adhered to plastic despite thorough washing. Only the bands that were different in respective test and control sample (mobbed red) were excised and analysed by MS. M denotes lanes loaded with molecular mass standards. Molecular mass is given in kDa.
Fig 2 Analysis of NPs protein corona using SDS-PAGE. PAA (A) and silica (B) NPs were dispersed in different media (dH2O, NaCl, PBS, RPMI) and incubated for 1 h in 10% or 100% FBS. Samples denoted with (+) contained PAA (A) or silica (B) NPs. In samples without NPs (-) were the proteins which adhered to plastic despite thorough washing. Only the bands that were different in respective test and control sample (mobbed red) were excised and analysed by MS. M denotes lanes loaded with molecular mass standards. Molecular mass is given in kDa.CC BY · PubMed Central
Fig 3 Relative abundance of proteins identified in samples of PAA NPs dispersed in different media. PAA NPs were dispersed in different media and incubated for 1h in 10% or 100% FBS. Proteins were separated from NPs, analysed on SDS-PAGE and identified by MS. Spectral counts were used as a measure of individual protein in a sample. Legend is further explained in Table 3. The differences in protein corona composition of PAA NPs dispersed in PBS, NaCl and RPMI-1640 were significant compared to protein corona of PAA NPs dispersed in dH2O (p < 0.05).
Fig 3 Relative abundance of proteins identified in samples of PAA NPs dispersed in different media. PAA NPs were dispersed in different media and incubated for 1h in 10% or 100% FBS. Proteins were separated from NPs, analysed on SDS-PAGE and identified by MS. Spectral counts were used as a measure of individual protein in a sample. Legend is further explained in Table 3. The differences in protein corona composition of PAA NPs dispersed in PBS, NaCl and RPMI-1640 were significant compared to protein corona of PAA NPs dispersed in dH2O (p < 0.05).CC BY · PubMed Central
Fig 4 Relative abundance of proteins identified in samples of silica NPs dispersed in different media. Silica NPs were dispersed in different media and incubated for 1h in 10% or 100% FBS. Proteins were separated from NPs, analysed on SDS-PAGE and identified by MS. Spectral counts were used as a measure of individual protein in a sample. Legend is further explained in Table 3. The differences in protein corona composition of silica NPs dispersed in PBS, NaCl and RPMI-1640 were significant compared to protein corona of silica NPs dispersed in dH2O (p < 0.05).
Fig 4 Relative abundance of proteins identified in samples of silica NPs dispersed in different media. Silica NPs were dispersed in different media and incubated for 1h in 10% or 100% FBS. Proteins were separated from NPs, analysed on SDS-PAGE and identified by MS. Spectral counts were used as a measure of individual protein in a sample. Legend is further explained in Table 3. The differences in protein corona composition of silica NPs dispersed in PBS, NaCl and RPMI-1640 were significant compared to protein corona of silica NPs dispersed in dH2O (p < 0.05).CC BY · PubMed Central
Fig 5 NPs protein coronas by theoretical pI of their constituting proteins. PAA (A, B) and silica (C, D) NPs were dispersed in different media and incubated for 1 h in 10% (A, C) or 100% (B, D) FBS. Proteins constituting NPs coronas were separated by SDS-PAGE and identified by MS. Theoretical pIs were calculated using Compute pI/Mw tool from the ExPASy portal.
Fig 5 NPs protein coronas by theoretical pI of their constituting proteins. PAA (A, B) and silica (C, D) NPs were dispersed in different media and incubated for 1 h in 10% (A, C) or 100% (B, D) FBS. Proteins constituting NPs coronas were separated by SDS-PAGE and identified by MS. Theoretical pIs were calculated using Compute pI/Mw tool from the ExPASy portal.CC BY · PubMed Central
Fig 6 NPs protein coronas by Gene Ontology (GO) biological process of constituting proteins. PAA (A, B) and silica (C, D) NPs were dispersed in different media and incubated for 1h in 10% (A, C) or 100% (B, D) FBS. Proteins forming corona of NPs were separated by SDS-PAGE and characterized by MS. GO information was acquired from the UniProt database.
Fig 6 NPs protein coronas by Gene Ontology (GO) biological process of constituting proteins. PAA (A, B) and silica (C, D) NPs were dispersed in different media and incubated for 1h in 10% (A, C) or 100% (B, D) FBS. Proteins forming corona of NPs were separated by SDS-PAGE and characterized by MS. GO information was acquired from the UniProt database.CC BY · PubMed Central

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Metadata

Authors
Strojan, K., Leonardi, A., Bregar, V. B., Križaj, I., Svete, J., & Pavlin, M.
Venue
PLOS ONE
Publisher
Public Library of Science (PLoS)
Published
2017-01-04
Volume
12
Issue
1
Pages
e0169552
ISSN
1932-6203
DOI
10.1371/journal.pone.0169552
PMC
PMC5215476
Cited by
131
References
3
Language
en
Licence
creativecommons.org/licenses/by/4.0/ (vor)
Funders
Javna Agencija za Raziskovalno Dejavnost RS (J7-7424, J3-6794, J2-6758, P1-0055, P1-0207, Young Researcher Program)

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Strojan, K., Leonardi, A., Bregar, V. B., Križaj, I., Svete, J., & Pavlin, M. (2017). Dispersion of Nanoparticles in Different Media Importantly Determines the Composition of Their Protein Corona. PLOS ONE. https://doi.org/10.1371/journal.pone.0169552