2D-electrophoresis and multiplex immunoassay proteomic analysis of different body fluids and cellular components reveal known and novel markers for extended fasting

Freek G Bouwman, Baukje De Roos, Isabel Rubio-Aliaga, L Katie Crosley, Susan J Duthie, Claus Mayer, Graham Horgan, Abigael C Polley, Carolin Heim, Susan LM Coort, Chris T Evelo, Francis Mulholland, Ian T Johnson, Ruan M Elliott, Hannelore Daniel, Edwin CM Mariman

    Research output: Contribution to journalArticle

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    Abstract

    Background

    Proteomic technologies applied for profiling human biofluids and blood cells are considered to reveal new biomarkers of exposure or provide insights into novel mechanisms of adaptation.

    Methods

    Both a non-targeted (classical 2D-electrophoresis combined with mass spectrometry) as well as a targeted proteomic approach (multiplex immunoassay) were applied to investigate how fasting for 36 h, as compared to 12 h, affects the proteome of platelets, peripheral blood mononuclear cells (PBMC), plasma, urine and saliva collected from ten healthy volunteers.

    Results

    Between-subject variability was highest in the plasma proteome and lowest in the PBMC proteome. Random Forests analysis performed on the entire dataset revealed that changes in the level of the RhoGDI2 protein in PBMC and plasma ApoA4 levels were the two most obvious biomarkers of an extended fasting. Random Forests (RF) analysis of the multiplex immunoassay data revealed leptin and MMP-3 as biomarkers for extended fasting. However, high between-subject variability may have masked the extended fasting effects in the proteome of the biofluids and blood cells.

    Conclusions

    Identification of significantly changed proteins in biofluids and blood cells using a non-targeted approach, together with the outcome of targeted analysis revealed both known and novel markers for a 36 h fasting period, including the cellular proteins RhoGDI2 and CLIC1, and plasma proteins ApoA4, leptin and MMP-3. The PBMC proteome exhibited the lowest between-subject variability and therefore these cells appear to represent the best biosamples for biomarker discovery in human nutrigenomics.
    Original languageEnglish
    Article number24
    Number of pages12
    JournalBMC Medical Genomics
    Volume4
    DOIs
    Publication statusPublished - 25 Mar 2011

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    Body Fluids
    Immunoassay
    Proteomics
    Electrophoresis
    Fasting
    Blood Cells
    Proteome
    Biomarkers
    Leptin
    Matrix Metalloproteinases
    Nutrigenomics
    Proteins
    Saliva
    Blood Proteins
    Mass Spectrometry
    Healthy Volunteers
    Blood Platelets
    Urine
    Technology

    Cite this

    2D-electrophoresis and multiplex immunoassay proteomic analysis of different body fluids and cellular components reveal known and novel markers for extended fasting. / Bouwman, Freek G; De Roos, Baukje; Rubio-Aliaga, Isabel; Crosley, L Katie; Duthie, Susan J; Mayer, Claus; Horgan, Graham; Polley, Abigael C; Heim, Carolin; Coort, Susan LM; Evelo, Chris T; Mulholland, Francis; Johnson, Ian T; Elliott, Ruan M; Daniel, Hannelore; Mariman, Edwin CM.

    In: BMC Medical Genomics, Vol. 4, 24, 25.03.2011.

    Research output: Contribution to journalArticle

    Bouwman, FG, De Roos, B, Rubio-Aliaga, I, Crosley, LK, Duthie, SJ, Mayer, C, Horgan, G, Polley, AC, Heim, C, Coort, SLM, Evelo, CT, Mulholland, F, Johnson, IT, Elliott, RM, Daniel, H & Mariman, ECM 2011, '2D-electrophoresis and multiplex immunoassay proteomic analysis of different body fluids and cellular components reveal known and novel markers for extended fasting', BMC Medical Genomics, vol. 4, 24. https://doi.org/10.1186/1755-8794-4-24
    Bouwman, Freek G ; De Roos, Baukje ; Rubio-Aliaga, Isabel ; Crosley, L Katie ; Duthie, Susan J ; Mayer, Claus ; Horgan, Graham ; Polley, Abigael C ; Heim, Carolin ; Coort, Susan LM ; Evelo, Chris T ; Mulholland, Francis ; Johnson, Ian T ; Elliott, Ruan M ; Daniel, Hannelore ; Mariman, Edwin CM. / 2D-electrophoresis and multiplex immunoassay proteomic analysis of different body fluids and cellular components reveal known and novel markers for extended fasting. In: BMC Medical Genomics. 2011 ; Vol. 4.
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    abstract = "BackgroundProteomic technologies applied for profiling human biofluids and blood cells are considered to reveal new biomarkers of exposure or provide insights into novel mechanisms of adaptation.MethodsBoth a non-targeted (classical 2D-electrophoresis combined with mass spectrometry) as well as a targeted proteomic approach (multiplex immunoassay) were applied to investigate how fasting for 36 h, as compared to 12 h, affects the proteome of platelets, peripheral blood mononuclear cells (PBMC), plasma, urine and saliva collected from ten healthy volunteers.ResultsBetween-subject variability was highest in the plasma proteome and lowest in the PBMC proteome. Random Forests analysis performed on the entire dataset revealed that changes in the level of the RhoGDI2 protein in PBMC and plasma ApoA4 levels were the two most obvious biomarkers of an extended fasting. Random Forests (RF) analysis of the multiplex immunoassay data revealed leptin and MMP-3 as biomarkers for extended fasting. However, high between-subject variability may have masked the extended fasting effects in the proteome of the biofluids and blood cells.ConclusionsIdentification of significantly changed proteins in biofluids and blood cells using a non-targeted approach, together with the outcome of targeted analysis revealed both known and novel markers for a 36 h fasting period, including the cellular proteins RhoGDI2 and CLIC1, and plasma proteins ApoA4, leptin and MMP-3. The PBMC proteome exhibited the lowest between-subject variability and therefore these cells appear to represent the best biosamples for biomarker discovery in human nutrigenomics.",
    author = "Bouwman, {Freek G} and {De Roos}, Baukje and Isabel Rubio-Aliaga and Crosley, {L Katie} and Duthie, {Susan J} and Claus Mayer and Graham Horgan and Polley, {Abigael C} and Carolin Heim and Coort, {Susan LM} and Evelo, {Chris T} and Francis Mulholland and Johnson, {Ian T} and Elliott, {Ruan M} and Hannelore Daniel and Mariman, {Edwin CM}",
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    AU - Bouwman, Freek G

    AU - De Roos, Baukje

    AU - Rubio-Aliaga, Isabel

    AU - Crosley, L Katie

    AU - Duthie, Susan J

    AU - Mayer, Claus

    AU - Horgan, Graham

    AU - Polley, Abigael C

    AU - Heim, Carolin

    AU - Coort, Susan LM

    AU - Evelo, Chris T

    AU - Mulholland, Francis

    AU - Johnson, Ian T

    AU - Elliott, Ruan M

    AU - Daniel, Hannelore

    AU - Mariman, Edwin CM

    PY - 2011/3/25

    Y1 - 2011/3/25

    N2 - BackgroundProteomic technologies applied for profiling human biofluids and blood cells are considered to reveal new biomarkers of exposure or provide insights into novel mechanisms of adaptation.MethodsBoth a non-targeted (classical 2D-electrophoresis combined with mass spectrometry) as well as a targeted proteomic approach (multiplex immunoassay) were applied to investigate how fasting for 36 h, as compared to 12 h, affects the proteome of platelets, peripheral blood mononuclear cells (PBMC), plasma, urine and saliva collected from ten healthy volunteers.ResultsBetween-subject variability was highest in the plasma proteome and lowest in the PBMC proteome. Random Forests analysis performed on the entire dataset revealed that changes in the level of the RhoGDI2 protein in PBMC and plasma ApoA4 levels were the two most obvious biomarkers of an extended fasting. Random Forests (RF) analysis of the multiplex immunoassay data revealed leptin and MMP-3 as biomarkers for extended fasting. However, high between-subject variability may have masked the extended fasting effects in the proteome of the biofluids and blood cells.ConclusionsIdentification of significantly changed proteins in biofluids and blood cells using a non-targeted approach, together with the outcome of targeted analysis revealed both known and novel markers for a 36 h fasting period, including the cellular proteins RhoGDI2 and CLIC1, and plasma proteins ApoA4, leptin and MMP-3. The PBMC proteome exhibited the lowest between-subject variability and therefore these cells appear to represent the best biosamples for biomarker discovery in human nutrigenomics.

    AB - BackgroundProteomic technologies applied for profiling human biofluids and blood cells are considered to reveal new biomarkers of exposure or provide insights into novel mechanisms of adaptation.MethodsBoth a non-targeted (classical 2D-electrophoresis combined with mass spectrometry) as well as a targeted proteomic approach (multiplex immunoassay) were applied to investigate how fasting for 36 h, as compared to 12 h, affects the proteome of platelets, peripheral blood mononuclear cells (PBMC), plasma, urine and saliva collected from ten healthy volunteers.ResultsBetween-subject variability was highest in the plasma proteome and lowest in the PBMC proteome. Random Forests analysis performed on the entire dataset revealed that changes in the level of the RhoGDI2 protein in PBMC and plasma ApoA4 levels were the two most obvious biomarkers of an extended fasting. Random Forests (RF) analysis of the multiplex immunoassay data revealed leptin and MMP-3 as biomarkers for extended fasting. However, high between-subject variability may have masked the extended fasting effects in the proteome of the biofluids and blood cells.ConclusionsIdentification of significantly changed proteins in biofluids and blood cells using a non-targeted approach, together with the outcome of targeted analysis revealed both known and novel markers for a 36 h fasting period, including the cellular proteins RhoGDI2 and CLIC1, and plasma proteins ApoA4, leptin and MMP-3. The PBMC proteome exhibited the lowest between-subject variability and therefore these cells appear to represent the best biosamples for biomarker discovery in human nutrigenomics.

    U2 - 10.1186/1755-8794-4-24

    DO - 10.1186/1755-8794-4-24

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    VL - 4

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    SN - 1755-8794

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    ER -