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Novel applications of mass spectrome...

Ngounou Wetie, Armand Gatien.

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Novel applications of mass spectrometry-based proteomics for biomarker discovery of autism spectrum disorder (ASD) and investigation of protein post-translational modifications (PTMs).

Record Type:	Electronic resources : Monograph/item
Title/Author:	Novel applications of mass spectrometry-based proteomics for biomarker discovery of autism spectrum disorder (ASD) and investigation of protein post-translational modifications (PTMs)./
Author:	Ngounou Wetie, Armand Gatien.
Description:	193 p.
Notes:	Source: Dissertation Abstracts International, Volume: 76-09(E), Section: B.
Contained By:	Dissertation Abstracts International76-09B(E).
Subject:	Chemistry. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3700332
ISBN:	9781321702347

Novel applications of mass spectrometry-based proteomics for biomarker discovery of autism spectrum disorder (ASD) and investigation of protein post-translational modifications (PTMs).
Ngounou Wetie, Armand Gatien.

Novel applications of mass spectrometry-based proteomics for biomarker discovery of autism spectrum disorder (ASD) and investigation of protein post-translational modifications (PTMs). - 193 p.

Source: Dissertation Abstracts International, Volume: 76-09(E), Section: B.

Thesis (Ph.D.)--Clarkson University, 2015.

The present work highlights the use of novel mass spectrometry-based methods for applications in biomedicine and protein chemistry. Generally, the work presents the development of novel methods to answer questions of unmet medical need and particularly pioneer the discovery of biological markers in neurodevelopmental disorders. In the first part, the focus is on the development of methods for the discovery of biologic markers of autism spectrum disorder (ASD) in different biological matrices such as blood serum and saliva. We made use of gel-based and nongel-based proteomic strategies. With regard to gel-based proteomics workflow, non-reducing Tricine-polyacrylamide gel electrophoresis (Tricine-PAGE) was used in combination with nano-liquid chromatography tandem mass spectrometry (nanoLC-MS/MS) for a pilot study of proteome profiling of the sera of children with ASD and typically developing control individuals. In another case, a two-dimensional gel electrophoresis (2-DE) approach that allowed for a better separation of complex salivary samples was adopted. Compared to the latter method, a nongel-based proteomic strategy (in-solution digestion) using a pooling strategy as commonly practiced in cancer biomarker studies was evaluated for ASD. All these studies provided strong putative biomarkers of ASD. We identified elevated levels of high density lipoproteins (HDL)-associated proteins paraoxonase 1, ApoA1 and ApoA4 pointing towards disruption of cholesterol metabolism in the serum of children with ASD compared to controls. The 2-DE approach identified a set of significantly dysregulated (up- and down-regulated) proteins, which could constitute a biomarker signature of ASD or ASD subtyping. There was some correlation of the result of this study with genes that were identified in previous studies as susceptible ASD risk factors. The same can be said of the pooling approach as well.

ISBN: 9781321702347Subjects--Topical Terms:

516420
Chemistry.

Novel applications of mass spectrometry-based proteomics for biomarker discovery of autism spectrum disorder (ASD) and investigation of protein post-translational modifications (PTMs).
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100 1 $a Ngounou Wetie, Armand Gatien. $3 3192938
245 1 0 $a Novel applications of mass spectrometry-based proteomics for biomarker discovery of autism spectrum disorder (ASD) and investigation of protein post-translational modifications (PTMs).
300 $a 193 p.
500 $a Source: Dissertation Abstracts International, Volume: 76-09(E), Section: B.
500 $a Adviser: Costel C. Darie.
502 $a Thesis (Ph.D.)--Clarkson University, 2015.
520 $a The present work highlights the use of novel mass spectrometry-based methods for applications in biomedicine and protein chemistry. Generally, the work presents the development of novel methods to answer questions of unmet medical need and particularly pioneer the discovery of biological markers in neurodevelopmental disorders. In the first part, the focus is on the development of methods for the discovery of biologic markers of autism spectrum disorder (ASD) in different biological matrices such as blood serum and saliva. We made use of gel-based and nongel-based proteomic strategies. With regard to gel-based proteomics workflow, non-reducing Tricine-polyacrylamide gel electrophoresis (Tricine-PAGE) was used in combination with nano-liquid chromatography tandem mass spectrometry (nanoLC-MS/MS) for a pilot study of proteome profiling of the sera of children with ASD and typically developing control individuals. In another case, a two-dimensional gel electrophoresis (2-DE) approach that allowed for a better separation of complex salivary samples was adopted. Compared to the latter method, a nongel-based proteomic strategy (in-solution digestion) using a pooling strategy as commonly practiced in cancer biomarker studies was evaluated for ASD. All these studies provided strong putative biomarkers of ASD. We identified elevated levels of high density lipoproteins (HDL)-associated proteins paraoxonase 1, ApoA1 and ApoA4 pointing towards disruption of cholesterol metabolism in the serum of children with ASD compared to controls. The 2-DE approach identified a set of significantly dysregulated (up- and down-regulated) proteins, which could constitute a biomarker signature of ASD or ASD subtyping. There was some correlation of the result of this study with genes that were identified in previous studies as susceptible ASD risk factors. The same can be said of the pooling approach as well.
520 $a In the second part of this thesis, we focus on the investigation of protein posttranslational modifications (PTMs), specifically on glycosylation and disulfide bridges. These PTMs affect protein stability, structure and function and therefore constitute a routine analysis of protein therapeutics. Two simple workflows of how to uncover these PTMs are shown. For glycosylation, identification of N-glycosylation sites of a recombinant protein using a Trypsin/AspN double digestion after treatment with PNGaseF revealing that the inclusion of potential glycosylation sites in a fusion protein can affect known and expected sites in the protein; a finding with significant consequence for the production of therapeutic glycoproteins. In the case of disulfide connectivities, the reference protein haptoglobin which can serve as a marker for disease state was exploited to provide the proof-of-concept of revealing disulfide connections using non-reducing SDS-PAGE in combination with western blotting (WB).
520 $a Finally, the last project was in the field of cancer biology focusing on a novel protein termed tumor differentiation factor (TDF) with promising differentiation activity specifically on breast and prostate cancer cells. In this part, efforts aiming at the characterization of the tdf gene in rat brains using in situ hybridization are shown and accord with previous expression work of the protein carried out in our group. The presence of the protein and its expression were also investigated in zebrafish for comparison with results obtained in rat and to get some clues with regard to its function. There are still many unknowns on TDF such as its native purification or the determination of its full cDNA sequence before its function and mode of action can be fully understood.
590 $a School code: 0049.
650 4 $a Chemistry. $3 516420
650 4 $a Biochemistry. $3 518028
650 4 $a Analytical chemistry. $3 3168300
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710 2 $a Clarkson University. $b Chemistry. $3 3177158
773 0 $t Dissertation Abstracts International $g 76-09B(E).
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791 $a Ph.D.
792 $a 2015
793 $a English
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3700332