Rapid discovery of putative protein biomarkers of traumatic brain injury by SDS-PAGE-capillary liquid chromatography-tandem mass spectrometry

William E. Haskins; Firas H. Kobeissy; Regina A. Wolper; Andrew K. Ottens; Jason W. Kitlen; Scott H. McClung; Barbara E. O'Steen; Marjorie M. Chow; Jose A. Pineda; Nancy D. Denslow; Ronald L. Hayes; Kevin K.W. Wang

doi:10.1089/neu.2005.22.629

Rapid discovery of putative protein biomarkers of traumatic brain injury by SDS-PAGE-capillary liquid chromatography-tandem mass spectrometry

William E. Haskins, Firas H. Kobeissy, Regina A. Wolper, Andrew K. Ottens, Jason W. Kitlen, Scott H. McClung, Barbara E. O'Steen, Marjorie M. Chow, Jose A. Pineda, Nancy D. Denslow, Ronald L. Hayes, Kevin K.W. Wang

Division of Critical Care Medicine

Research output: Contribution to journal › Article › peer-review

54 Scopus citations

Abstract

We report the rapid discovery of putative protein biomarkers of traumatic brain injury (TBI) by SDS-PAGE-capillary liquid chromatography-tandem mass spectrometry (SDS-PAGE-Capillary LC-MS²). Ipsilateral hippocampus (IH) samples were collected from naive rats and rats subjected to controlled cortical impact (a rodent model of TBI). Protein database searching with 15,558 uninterpreted MS² spectra, collected in 3 days via data-dependent capillary LC-MS² of pooled cyanine dye-labeled samples separated by SDS-PAGE, identified more than 306 unique proteins. Differential proteomic analysis revealed differences in protein sequence coverage for 170 mammalian proteins (57 in naive only, 74 in injured only, and 39 of 64 in both), suggesting these are putative biomarkers of TBI. Confidence in our results was obtained by the presence of several known biomarkers of TBI (including αII-spectrin, brain creatine kinase, and neuron-specific enolase) in our data set. These results show that SDS-PAGE prior to in vitro proteolysis and capillary LC-MS² is a promising strategy for the rapid discovery of putative protein biomarkers associated with a specific physiological state (i.e., TBI) without a priori knowledge of the molecules involved.

Original language	English
Pages (from-to)	629-644
Number of pages	16
Journal	Journal of neurotrauma
Volume	22
Issue number	6
DOIs	https://doi.org/10.1089/neu.2005.22.629
State	Published - Jun 2005

Keywords

Controlled cortical impact (CCI)
Differential in-gel electrophoresis (DIGE)
Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)
Tandem mass spectometry (MS)
Traumatic brain injury (TBI)

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Haskins, W. E., Kobeissy, F. H., Wolper, R. A., Ottens, A. K., Kitlen, J. W., McClung, S. H., O'Steen, B. E., Chow, M. M., Pineda, J. A., Denslow, N. D., Hayes, R. L., & Wang, K. K. W. (2005). Rapid discovery of putative protein biomarkers of traumatic brain injury by SDS-PAGE-capillary liquid chromatography-tandem mass spectrometry. Journal of neurotrauma, 22(6), 629-644. https://doi.org/10.1089/neu.2005.22.629

@article{bae5f06707ba4db18eca231b45d036ab,

title = "Rapid discovery of putative protein biomarkers of traumatic brain injury by SDS-PAGE-capillary liquid chromatography-tandem mass spectrometry",

abstract = "We report the rapid discovery of putative protein biomarkers of traumatic brain injury (TBI) by SDS-PAGE-capillary liquid chromatography-tandem mass spectrometry (SDS-PAGE-Capillary LC-MS2). Ipsilateral hippocampus (IH) samples were collected from naive rats and rats subjected to controlled cortical impact (a rodent model of TBI). Protein database searching with 15,558 uninterpreted MS2 spectra, collected in 3 days via data-dependent capillary LC-MS2 of pooled cyanine dye-labeled samples separated by SDS-PAGE, identified more than 306 unique proteins. Differential proteomic analysis revealed differences in protein sequence coverage for 170 mammalian proteins (57 in naive only, 74 in injured only, and 39 of 64 in both), suggesting these are putative biomarkers of TBI. Confidence in our results was obtained by the presence of several known biomarkers of TBI (including αII-spectrin, brain creatine kinase, and neuron-specific enolase) in our data set. These results show that SDS-PAGE prior to in vitro proteolysis and capillary LC-MS2 is a promising strategy for the rapid discovery of putative protein biomarkers associated with a specific physiological state (i.e., TBI) without a priori knowledge of the molecules involved.",

keywords = "Controlled cortical impact (CCI), Differential in-gel electrophoresis (DIGE), Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Tandem mass spectometry (MS), Traumatic brain injury (TBI)",

author = "Haskins, {William E.} and Kobeissy, {Firas H.} and Wolper, {Regina A.} and Ottens, {Andrew K.} and Kitlen, {Jason W.} and McClung, {Scott H.} and O'Steen, {Barbara E.} and Chow, {Marjorie M.} and Pineda, {Jose A.} and Denslow, {Nancy D.} and Hayes, {Ronald L.} and Wang, {Kevin K.W.}",

year = "2005",

month = jun,

doi = "10.1089/neu.2005.22.629",

language = "English",

volume = "22",

pages = "629--644",

journal = "Journal of neurotrauma",

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Haskins, WE, Kobeissy, FH, Wolper, RA, Ottens, AK, Kitlen, JW, McClung, SH, O'Steen, BE, Chow, MM, Pineda, JA, Denslow, ND, Hayes, RL & Wang, KKW 2005, 'Rapid discovery of putative protein biomarkers of traumatic brain injury by SDS-PAGE-capillary liquid chromatography-tandem mass spectrometry', Journal of neurotrauma, vol. 22, no. 6, pp. 629-644. https://doi.org/10.1089/neu.2005.22.629

TY - JOUR

T1 - Rapid discovery of putative protein biomarkers of traumatic brain injury by SDS-PAGE-capillary liquid chromatography-tandem mass spectrometry

AU - Haskins, William E.

AU - Kobeissy, Firas H.

AU - Wolper, Regina A.

AU - Ottens, Andrew K.

AU - Kitlen, Jason W.

AU - McClung, Scott H.

AU - O'Steen, Barbara E.

AU - Chow, Marjorie M.

AU - Pineda, Jose A.

AU - Denslow, Nancy D.

AU - Hayes, Ronald L.

AU - Wang, Kevin K.W.

PY - 2005/6

Y1 - 2005/6

N2 - We report the rapid discovery of putative protein biomarkers of traumatic brain injury (TBI) by SDS-PAGE-capillary liquid chromatography-tandem mass spectrometry (SDS-PAGE-Capillary LC-MS2). Ipsilateral hippocampus (IH) samples were collected from naive rats and rats subjected to controlled cortical impact (a rodent model of TBI). Protein database searching with 15,558 uninterpreted MS2 spectra, collected in 3 days via data-dependent capillary LC-MS2 of pooled cyanine dye-labeled samples separated by SDS-PAGE, identified more than 306 unique proteins. Differential proteomic analysis revealed differences in protein sequence coverage for 170 mammalian proteins (57 in naive only, 74 in injured only, and 39 of 64 in both), suggesting these are putative biomarkers of TBI. Confidence in our results was obtained by the presence of several known biomarkers of TBI (including αII-spectrin, brain creatine kinase, and neuron-specific enolase) in our data set. These results show that SDS-PAGE prior to in vitro proteolysis and capillary LC-MS2 is a promising strategy for the rapid discovery of putative protein biomarkers associated with a specific physiological state (i.e., TBI) without a priori knowledge of the molecules involved.

AB - We report the rapid discovery of putative protein biomarkers of traumatic brain injury (TBI) by SDS-PAGE-capillary liquid chromatography-tandem mass spectrometry (SDS-PAGE-Capillary LC-MS2). Ipsilateral hippocampus (IH) samples were collected from naive rats and rats subjected to controlled cortical impact (a rodent model of TBI). Protein database searching with 15,558 uninterpreted MS2 spectra, collected in 3 days via data-dependent capillary LC-MS2 of pooled cyanine dye-labeled samples separated by SDS-PAGE, identified more than 306 unique proteins. Differential proteomic analysis revealed differences in protein sequence coverage for 170 mammalian proteins (57 in naive only, 74 in injured only, and 39 of 64 in both), suggesting these are putative biomarkers of TBI. Confidence in our results was obtained by the presence of several known biomarkers of TBI (including αII-spectrin, brain creatine kinase, and neuron-specific enolase) in our data set. These results show that SDS-PAGE prior to in vitro proteolysis and capillary LC-MS2 is a promising strategy for the rapid discovery of putative protein biomarkers associated with a specific physiological state (i.e., TBI) without a priori knowledge of the molecules involved.

KW - Controlled cortical impact (CCI)

KW - Differential in-gel electrophoresis (DIGE)

KW - Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)

KW - Tandem mass spectometry (MS)

KW - Traumatic brain injury (TBI)

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U2 - 10.1089/neu.2005.22.629

DO - 10.1089/neu.2005.22.629

M3 - Article

C2 - 15941373

AN - SCOPUS:20544442753

SN - 0897-7151

VL - 22

SP - 629

EP - 644

JO - Journal of neurotrauma

JF - Journal of neurotrauma

IS - 6

ER -

Rapid discovery of putative protein biomarkers of traumatic brain injury by SDS-PAGE-capillary liquid chromatography-tandem mass spectrometry

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Keywords

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