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Biomarker Discovery for Alzheimer’s Disease Using NAPPA and In Vivo Crystallization in Baculovirus-Infected Insect Cells for Structural Biology

Description

Proteins are a large collection of biomolecules that orchestrate the vital

cellular processes of life. The last decade has witnessed dramatic advances in the

field of proteomics, which broadly include characterizing the

Proteins are a large collection of biomolecules that orchestrate the vital

cellular processes of life. The last decade has witnessed dramatic advances in the

field of proteomics, which broadly include characterizing the composition, structure,

functions, interactions, and modifications of numerous proteins in biological systems,

and elucidating how the miscellaneous components collectively contribute to the

phenotypes associated with various disorders. Such large-scale proteomics studies

have steadily gained momentum with the evolution of diverse high-throughput

technologies. This work illustrates the development of novel high-throughput

proteomics platforms and their applications in translational and structural biology. In

Chapter 1, nucleic acid programmable protein arrays displaying the human

proteomes were applied to immunoprofiling of paired serum and cerebrospinal fluid

samples from patients with Alzheimer’s disease. This high-throughput

immunoproteomic approach allows us to investigate the global antibody responses

associated with Alzheimer’s disease and potentially identify the diagnostic

autoantibody biomarkers. In Chapter 2, a versatile proteomic pipeline based on the

baculovirus-insect cell expression system was established to enable high-throughput

gene cloning, protein production, in vivo crystallization and sample preparation for Xray diffraction. In conjunction with the advanced crystallography methods, this endto-end pipeline promises to substantially facilitate the protein structural

determination. In Chapter 3, modified nucleic acid programmable protein arrays

were developed and used for probing protein-protein interactions at the proteome

level. From the perspective of biomarker discovery, structural proteomics, and

protein interaction networks, this work demonstrated the power of high-throughput

proteomics technologies in myriad applications for proteome-scale structural,

functional, and biomedical research.

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Date Created
  • 2020