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cell-free-protein-expression-hibit.md

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1# Cell Free Protein Expression with HiBiT Quantification

3**URL:** https://cloud.ginkgo.bio/protocols/cell-free-protein-expression-hibit

4**Service terms:** https://cloud.ginkgo.bio/terms/cfps-hibit

5**Reviewed:** 2026-09-30; prices and turnaround below are catalog estimates.

6**Status:** Ginkgo Certified

7**Price:** $39/sample

8**Turnaround:** up to 11 days

9**Throughput:** 384-well format

Cell Free Protein Expression with HiBiT Quantification

URL: https://cloud.ginkgo.bio/protocols/cell-free-protein-expression-hibit Service terms: https://cloud.ginkgo.bio/terms/cfps-hibit Reviewed: 2026-09-30; prices and turnaround below are catalog estimates. Status: Ginkgo Certified Price: $39/sample Turnaround: up to 11 days Throughput: 384-well format

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11## Overview

13Fastest 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.

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.

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15## Input

17- **Sequence input:** The protocol page links a CFPS `.xlsx` template for linear DNA; service terms describe protein or DNA in CSV. Confirm the downloaded schema and synthesis format (linear DNA or clonal plasmid).

18- **HiBiT Tag Orientation:** N-terminal or C-terminal fusion

19- **Linker Sequence:** If unspecified, Ginkgo uses a standard GGGS linker

Input

Sequence input: The protocol page links a CFPS .xlsx template for linear DNA; service terms describe protein or DNA in CSV. Confirm the downloaded schema and synthesis format (linear DNA or clonal plasmid).
HiBiT Tag Orientation: N-terminal or C-terminal fusion
Linker Sequence: If unspecified, Ginkgo uses a standard GGGS linker

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21## Output

23- **Expression Detection:** Confirmation of target protein expression via background-subtracted luminescence (bcRLU) relative to controls

24- **Relative Yield Quantification:** Target protein concentration (reported in nM), interpolated from a standard curve

25- **Assay Quality Metrics:** Z-prime scores, replicate CV%, and matrix-effect controls

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

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27## Automated Workflow