Stable Cell Line Development Service – Constitutive Expression
Our proprietary ExoIN technology is a powerful and reliable platform for the development of customized assay cell lines with stable and homogeneous target expression. Using this technology, we are able to generate your desired mammalian cell line to support your specific project research requirements within weeks instead of months. All we need is your target gene sequence to start your project.
trenzyme’s ExoIN Technology – Modules & Explanation
The ExoIN technology links target protein expression to the expression of a resistance marker. The target, the selection marker and the ExoIN module are transcribed together as a single mRNA transcript. Subsequent efficient cotranslational cleavage at the ribosome yields the desired target protein unmodified and functional in a 1-to-1 stoichiometry with the selection marker. This means that once selected, the stable cell pool transcribes the target protein almost homogeneously (e.g. reference protein expression GFP > 90%).
This unique technology allows an easy and highly efficient selection of homogeneous cell populations – in consequence, time consuming and expensive single cell cloning is no longer necessary for many applications.
Main Advantages of trenzyme’s ExoIN Technology
- Generation of ready-to-use stable assay cell lines within weeks – no single cell cloning needed depending on target and parental cell line.
- Fast feasibility analysis – cell growth under selection conditions correlates with constitutive target expression.
- The appearance of false-positive cells (resistant to selection marker but without target expression) can be minimized by our ExoIN technology.
- ExoIN technology is applicable to any cell line and all classes of target proteins.
- Due to perfect co-translational cleavage the target remains unmodified and functional. No uncut fusion proteins detectable.
- The expression level remains stable as long as the selection pressure is constant.
- Three different selection markers (puromycin, hygromycin and zeocin) allowing simultaneous selection up to three targets.
„We were astonished by trenzyme’s recently delivered custom ExoIN cell lines. Even „difficult-to-express“ constructs show stable and robust expression. I highly recommend trenzyme’s cell line development service.“
Process of Stable Cell Line Development
Stable Cell Line Development Service - Overview
Quality Control Assays
To ensure highest quality and validity for our final products, we offer a broad range of quality control assays for our cell line development services.
- Mycoplasma check provided by our partner Eurofins Genomics
- Viability assay
- Assessment of target expression: mRNA (via qPCR) or protein (via FACS or Western Blot)
- Cell based assay for validation: e.g. Calcium flux assay, apoptosis assay, reporter assays or phosphorylation assessment
- Microscopic analysis
- Sterility assay
- Cell line authentication by STR profiling
- Copy number determination
Further Options for Cell Line Development
- Provide your preferred parental cell line and/or your expression system of choice and we will generate the recombinant cell line according to your needs.
- Inducible target expression
Constitutive Expression FAQs
How many vials and how many cells per vial are delivered?
Our standard delivery comprises 3 cryovials per stable pool and 3 cryovials per each of the 3 best clones with 1e6 cells per vial for adherent cells or up to 1e7 cells for suspensions cells. Additional vials are possible on request.
Do royalties have to be paid?
The ExoIN cell lines are royalty free.
How many clones do you generate?
Our standard is to expand 30 and test up to 24 clones by the agreed method and deliver the best three clones according the analyses. If you have other requirements, we will discuss your needs and can adapt our standard service to a certain extent.
Dr. Antje Fuhrmann, PhD
Application & Sales Manager
We would be happy to provide you with support on your cell line research project. Let us know your questions and requests about our cell line services and/or processes – our scientific experts will reply shortly.
Get In Contact With Our Scientific Experts