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86 lines
2.6 KiB
Markdown
86 lines
2.6 KiB
Markdown
# Cell Free Protein Expression Optimization
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**URL:** https://cloud.ginkgo.bio/protocols/cell-free-protein-expression-optimization
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**Status:** Ginkgo Certified
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**Price:** $199/sample (default: $597 for 1 protein x 3 replicates = 3 samples)
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**Turnaround:** 6-11 days
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## Overview
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Design of Experiment (DoE) approach to expressing protein targets in a proprietary reconstituted E. coli transcription-translation system. Each construct is evaluated in up to 24 reaction conditions per protein, including target-specific additives such as chaperones, disulfide-bond enhancers, and cofactors. Designed for difficult-to-express proteins including membrane proteins and targets with disulfide or cofactor requirements.
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## Input
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- **DNA sequence** in `.fasta` format
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## Output
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- **Comparative Yield:** Titer data mapped across all tested variables (lysates, temps, additives)
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- **Purity Profiling:** Target protein vs. background impurities to find highest quality yield
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- **Optimal Conditions:** Overlaid electropherograms pinpointing the exact formulation for a given sequence
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## Automated Workflow
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### Phase 1 - Reagent Prep
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1. Retrieve plates from 4 deg C
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2. Thaw at room temperature
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3. PBS backfill
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### Phase 2 - CFPS Reaction Setup & Incubation
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1. Retrieve plates from 4 deg C
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2. Dispense lysate
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3. QC plate read
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4. Incubate (shaking or static, condition-dependent)
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### Phase 3 - Quantification Prep & Read
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1. Dispense PBS
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2. Unseal plate
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3. LabChip quantification
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4. Seal plate
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5. Store at 4 deg C
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## Protocol Parameters
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- Payloads & Reagents
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- Bravo Stamp
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- HiG Centrifuge
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- Incubation & Storage
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## Optimization Variables
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The DoE matrix can span up to 24 conditions per protein, varying:
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- **Lysate composition** (different E. coli extract formulations)
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- **Temperature** (incubation temperature profiles)
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- **Additives:**
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- Chaperones (for folding-challenged targets)
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- Disulfide-bond enhancers (for targets requiring disulfide bridges)
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- Cofactors (metal ions, coenzymes, prosthetic groups)
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- Other target-specific supplements
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## Ordering
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- **Number of Proteins:** configurable
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- **Number of Replicates:** configurable
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- **File Upload:** CSV, Excel, FASTA, TXT, PDF, ZIP
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- **Additional Details:** free-text field for special requirements
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## Certification Milestones
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- Dry Run Complete
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- Wet Run Complete
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- Biovalidation Complete
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- App Note Complete
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## Use Cases
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- Optimizing expression of difficult-to-express proteins
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- Membrane protein expression screening
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- Identifying optimal conditions for disulfide-bonded proteins
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- Cofactor-dependent protein expression
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- Systematic exploration of expression parameter space
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- Finding the best formulation before scaling up production
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