Arnaud Monnard - rna.uzh.chffffffff-b34e-2810-ffff... · Cell Free • PISA • NAPPA • In Situ...
Transcript of Arnaud Monnard - rna.uzh.chffffffff-b34e-2810-ffff... · Cell Free • PISA • NAPPA • In Situ...
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Protein Microarrays Journal Club
Arnaud Monnard
August 26th 2014 Aguzzi lab
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Outline Manufacturing of microarrays • Contact Printing
• Pin Printing • Microstamping • Flow Printing
• Lithography • Photolithography • Electron Beam • Dip Pen Nanolithography
• Non Contact Printing • Thermal • Piezo • Valve
• Cell Free • PISA • NAPPA • In Situ Puromycin Capture
Publications
• Proof of Concepts • MacBeath et al. Science 2000 • Chen et al. BioTechniques 2006
• Applications • Jones et al. Nature 2006 • Mok et al. Nature Protocols 2009 • Gagni et al. Biosensors and
Bioelectronics 2013
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Introduction
• First antibody array in 1983 followed by developement in the late 90’s
• Use principles and technologies of DNA microarrays – Minimal sample consumption – High throughput
• Bypasses the limitation of DNA microarrays
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Protein Microarray Manufacturing Technologies
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Contact Printing
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Pin Printing
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Microstamping
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Flow Printing
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Lithography
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Photolithography
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Electron Beam
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Dip Pen Nanolithography
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Non-Contact Printing
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Non contact Printing
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Cell Free Systems
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Protein In Situ Array (PISA)
No DNA immobilization
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NAPPA
DNA immobilization
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In Situ Puromycin Capture
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Proof of Concept Protein-Protein interaction
No rapamycin
Single spot detection
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Proof of Concept Detection of kinase substrates Detection of small molecules targets
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Applications
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Binding Affinity
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Quantitative Interaction
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Quantitative Interaction
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System-level View
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Available Domains
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Yeast Kinase Identification
Negative control
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Capture Ab selection
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AB 42 Detection
Artificial CSF
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Signal to Noise ratio
Human CSF
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Conclusions • 15 years after the first landmark papers and proof
of concepts the technique is limited to a few applications
• Use of arrays is discouraged by – The laborious preparation processes – Cost and equipment – Complexity of data generated
• Protein folding and missfolding remain a challenge and an obstacle for protein chip
• Protein chips are already available commercially, but major publications validating their use are still missing
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Questions?