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