Table 2. Technology Development Roadmap for Complex Isolation

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Table 2. Technology Development Roadmap for Complex Isolation (acronyms)

Technology Objectives	Research, Design, and Development	Demonstration: Pilots and Modular Deployment	Integration and Production Deployment	Facility Outputs
Develop high-throughput technologies for molecular machine isolation for full population of biomolecular complexes Soluble stable complexes Membrane-associated complexes Transient complexes	Define needs: Evaluation of commercial laboratory and LIMS resources, if available Protocol refinement, automation QA/QC Multistage isolation schemes using affinity reagents to minimize background interferences Microfluidic-based affinity reagent isolations to minimize sample size requirements Stabilization and cross-linking of less-stable and transient complexes In vivo validation approaches Develop: Continuous, automated processing Multiplexed pulldowns Novel affinity reagents and isolation schemes Develop: Solubilization of membrane-associated complexes Stabilization of complexes Develop stabilization and cross-linking	Pilot-scale isolation method: Scaleup from 100 assays per week to thousands per week Automated, continuous processing Assessment of bottlenecks, costs QA/QC Evaluation and incorporation of new technologies Methods for rapid elucidation of protein complex network linkage maps	Establishment of multiple parallel pipelines Evaluation and incorporation of new technologies	Complexes isolated to permit identification, imaging, and biophysical characterization Protocols Methods Databases and query tools

Technology Objectives

Research, Design, and Development

Demonstration:
Pilots and Modular Deployment

Integration and Production Deployment

Facility Outputs

Develop high-throughput technologies for molecular machine isolation for full population of biomolecular complexes

Soluble stable complexes

Membrane-associated complexes

Transient complexes

Define needs:

Evaluation of commercial laboratory and LIMS resources, if available
Protocol refinement, automation
QA/QC
Multistage isolation schemes using affinity reagents to minimize background interferences
Microfluidic-based affinity reagent isolations to minimize sample size requirements
Stabilization and cross-linking of less-stable and transient complexes
In vivo validation approaches

Develop:

Continuous, automated processing
Multiplexed pulldowns
Novel affinity reagents and isolation schemes

Develop:

Solubilization of membrane-associated complexes
Stabilization of complexes

Develop stabilization and cross-linking

Pilot-scale isolation method:

Scaleup from 100 assays per week to thousands per week
Automated, continuous processing
Assessment of bottlenecks, costs
QA/QC
Evaluation and incorporation of new technologies
Methods for rapid elucidation of protein complex network linkage maps

Establishment of multiple parallel pipelines

Evaluation and incorporation of new technologies

Complexes isolated to permit identification, imaging, and biophysical characterization

Protocols

Methods

Databases and query tools

To develop and incorporate the necessary technologies and methods into a high-throughput production environment, a phased process will be followed as described in this roadmap. The process includes research, design, and development; modular and pilot-scale deployment; and final integration and scaleup into operational procedures.

U.S. Department of Energy Office of Science

Genomics:GTL

Table 2. Technology Development Roadmap for Complex Isolation (acronyms)

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