Cell Free Protein Expression with HiBiT Quantification

URL: https://cloud.ginkgo.bio/protocols/cell-free-protein-expression-hibit Status: Ginkgo Certified Price: $39/sample Turnaround: up to 11 days Throughput: 384-well format

Overview

Fastest path from a sequence to a protein yield metric. Uses a proprietary reconstituted E. coli transcription-translation (CFPS) system; reactions complete in 4-16 hours and can yield up to 3 mg/mL of target protein. Expressed proteins are quantified directly from the crude reaction mixture (no purification) using the Promega Nano-Glo HiBiT Lytic Detection System. Designed for early-stage screening, novel construct evaluation, and rapid, data-driven triage before committing to optimization or purification.

Input

• DNA Input: Linear DNA sequence (.xlsx template)
• HiBiT Tag Orientation: N-terminal or C-terminal fusion
• Linker Sequence: If unspecified, Ginkgo uses a standard GGGS linker

Output

• Expression Detection: Confirmation of target protein expression via background-subtracted luminescence (bcRLU) relative to controls
• Relative Yield Quantification: Target protein concentration (reported in nM), interpolated from a standard curve
• Assay Quality Metrics: Z-prime scores, replicate CV%, and matrix-effect controls

Automated Workflow

Phase 1 - CFPS Reaction Setup & Incubation

1. Stamp DNA templates into CFPS reaction mix
2. Seal and incubate (4-16 h)

Phase 2 - Detection Prep

1. Add Nano-Glo HiBiT Lytic detection reagent

Phase 3 - Detection & Quantification

1. Incubate (ambistore)
2. Luminescence read (BMG PHERAstar)
3. Seal & store (Agilent PlateLoc)

Ordering

• Number of Proteins: configurable
• Number of Replicates: configurable
• File Upload: CSV, Excel, FASTA, TXT, PDF, ZIP
• Additional Details: free-text field for special requirements

Use Cases

• High-throughput expressibility screening without purification
• Rapid relative-yield comparison across construct variants
• Early triage before A280/LabChip purification tiers