Ginkgo’s CFPS Disulfide Premium kit is tailored for the production of proteins containing disulfide bonds. It enables the synthesis of functional disulfide bonded targets that can’t properly express with standard CFPS, including antibody fragments, multi-domain therapeutic enzymes, and more. The kit is an E. coli transcription/translation system optimized to produce proteins from a T7 RNA Polymerase promoter under redox conditions that facilitate disulfide bond (DSB) formation. Reactions consist of a bacterial cell lysate, a reagent mix containing all components required for protein production, and a DSB Booster that drives formation and correct pairing of DSBs in the target protein. The user must simply provide a DNA template that encodes the protein of interest under control of a T7 promoter. Reactions are assembled on ice, incubated at 30 °C for 4 - 18 hours, and can be analyzed in solution or after purification. The system has been optimized for proteins carrying 1 to 9 disulfide bonds and validated across protein classes spanning that range: Antibody fragments: VHHs and scFvs (1–2 disulfide bonds), expressed with >95% DSB confirmed via intact mass spectrometry) Reporter enzymes: Gaussia luciferase (5 disulfide bonds), supplied as the kit's positive control Multi-domain therapeutic enzymes: reteplase (9 disulfide bonds), a plasminogen activator that is notoriously difficult to express solubly in conventional E. coli systems Contents: DSB E. coli Lysate DSB Reagent Mix (3.3x) DSB Booster (20x) Control Gaussia Luciferase (GLuc) Plasmid (100 nM) Technical Information: Optimized for targets containing 1 to 9 disulfide bonds. Compatible with both plasmid DNA and linear DNA templates. Eliminate time-consuming cloning steps by using linear DNA directly. Expression is driven by a T7 RNA polymerase promoter. Reaction volumes can be scaled up or down proportionally from the standard recommended 20 uL setup. Includes a Gaussia luciferase (Gluc) positive control to verify successful CFPS. Incubation at 30 °C for 4 to 18 hours is ideal across various protein classes (although target-specific optimization may improve titers). Kit is stored at −80 °C. GLuc Control Plasmid at −20 °C. The kit ships on dry ice. This beta release is not compatible with downstream purification via His-tag (IMAC/Ni-NTA) affinity. Use an alternative affinity tag such as FLAG or Strep-II, or recover protein by size-exclusion or ion-exchange chromatography. This kit is for research use only and is not for use in diagnostic or therapeutic procedures. Quick Start Guide CFPS Disulfide User ManualControl Gaussia Luciferase (GLuc) Plasmid Sequence Terms of Use Safety Data Sheets: DSB Lysate SDS, DSB Reagent Mix SDS, DSB Booster SDS, Control GLuc Plasmid SDS Cell-Free Protein Synthesis kits are for research use only. Administration to human and other animals and use in the clinic, for diagnostic purposes, and other uses are prohibited. Do not use in foods or for domestic purposes. Do you have questions about our kit or want a consult on how to best use Ginkgo's CFPS kits? Are you interested in a custom order? Reach out to us at [email protected]
