FAQs

Yes, there is a specific project manager assigned to every project.

You will receive weekly updates on the current process status and timeline of your project by your assigned project manager.

Yes, we offer molecular biology services, both as addition to our cell line development and protein production services and as stand-alone service.

We discuss upfront the area you are most interested in and then propose the current options for you in detail.

The minimum duration of an internship is three months.

The timeline for the generation of a basic pool starts at 5 weeks and for clonal cell lines at 13 weeks.

Depending on the target and your specific requirements we can test the stable cell line for GOI expression by a variety of methods. You can choose from qPCR, Western blot, FACS analysis, ELISA, IF or cell-based assays.

The data sheet contains all relevant information regarding your cell line in a comprehensive and short version.

We will generate a detailed documentation containing all project results and protocols as well as a detailed list of materials and substances (including lot numbers). You will obtain a draft version of the report for review. The report will be finalized after receiving your amendments (one round).

We already worked with dozens of human (e.g. HT1080, Jurkat,…) and non-human cell lines (e.g. NIH3T3, PC12,…).

Routinely we apply flow cytometry based cell sorting or limiting dilution for single cell cloning. For difficult target cell lines we also offer colony picking in soft agar.

For ExoIN technology we can offer puromycine, hygromycine and zeocin as possible resistance markers.

You can choose between our in-house cell lines HEK293 and CHO-K1 or you can send us your parental cell line of choice.

The ExoIN technology links target protein expression to the expression of a resistance marker on protein level. The target, the selection marker and the ExoIN module are transcribed together and initially a fusion protein is translated. This is cleaved co-translationally with highest efficiency, so your target is synthesized as free, non-modified protein.

No, there are no additional amino acids remaining as the tag gets completely cleaved off. The target is unmodified after cleavage.

No, if you provide your preferred parental cell line and/or your expression system of choice, we will generate the recombinant cell line according to your needs.

Our standard method is electroporation for the generation of stable assay cell lines, but lipofection or other chemical transfection methods are possible as well if required.

Yes, the data sheet contains a BSL classification, which depends on the target and parental cell line of choice (german Gen-TG).

Yes, we have a lot of experience with other expression systems. But we prefer our ExoIN system since it is the most powerful expression system with a lot of benefits.

Our standard promoter is CMV but other promoters like EF-1 are also available.

The ExoIN cell lines are royalty free.

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.

No, we offer only customized recombinant cell lines and actually no cell lines off the shelf.

The ExoIN tag is located at the N-terminus of the protein. Subsequent efficient cotranslational cleavage at the ribosome yields the desired free target protein unmodified and functional in a 1-to-1 stoichiometry with the selection marker.

We already worked with all different target classes: transmembrane receptors, channels & transporters, intracelllular proteins, secreted proteins and many other target classes.

No, if you provide your preferred parental cell line and your inducible expression system of choice, we will generate the recombinant cell line according to your needs.

The regulation of the target expression in the ExoINDUCE system is under control of a proprietary version of the tetracycline transactivator (rtTA). It is an optimized “Tet-on” system, meaning the expression of the target is going to be induced by addition of doxycycline to the media.

As a high standard productivity, we can reach up to 3g/l for mABs after cell line development. After process development, it can reach > 8 g/l.

Within the cell line generation at trenzyme, a R&D license is already included and has not to be purchased. For any commercial application later on, a commercial license is needed.

We can optionally offer a stability study and also a copy number determination, if desired.

We can additionally offer development of the Downstream Processing (DSP), protein purification from fed-batch production and a variety of protein analysis services (e.g. mass spectrometry, protein sequencing, HP-SEC) as well as metabolite analyses during fed-batch small scale production.

Our express version takes 16-20 weeks and includes two rounds of fed-batch-screening.

Depending on the target and your specific requirements we can analyze titer, cell growth properties, target gene copy number and stability as well as product quality.

Although we do have internal iPSC lines and validated differentiation protocols, we do not sell these iPSC lines or iPSC-derived progeny.

Yes, we do have internal differentiation protocols (e.g. neuronal, or hepatocyte protocols), which can be used in order to provide customized iPSC-derived cells. You will receive the generated and cryopreserved cells, as well as a comprehensive data sheet, however, no details regarding the differentiation protocol will be given.

We routinely perform immunofluorescence staining, FACs analysis and qPCR in order to characterize the stages of differentiation. For special read-outs, do not hesitate to contact our experts.

During the project, regular telephone calls, email correspondence and short reports ensure detailed exchange of information and results. Furthermore, technical discussions will help to implement key experiment layouts according to your needs.

In order to characterize iPSC lines for the expression of pluripotency markers, immunoflourescence staining and/or flow cytometry for key pluripotency markers OCT4, SOX2, SSEA-4, TRA-1-60, and TRA-1-81 will be performed. Furthermore, the ability to form all three germ layers will be investigated via directed differentiation towards endo-, meso-, and ectoderm (immunoflourescence staining, RT-qPCR).

In order to evaluate genomic stability, chromosomal abnormalities, and copy number variants, molecular karyotyping via Illumina BeadArray will be applied.

With our cell expansion service we can generate up to 1e9 cells in one batch.

Our scientific experts can develop assays based on basic cell lines, customized cell lines or iPSC-derived cell types. For this, we use our in-house differentiation protocols with cardiomyocytes, hepatocyte-like cells or neurons for your analysis. Alternatively, you can provide us with your protocol so that after the technology transfer, our experts perform the work using this protocol internally at trenzyme.

Yes, our assay experts are delighted to develop an assay according to your needs prior to compound analysis.

We measure small and medium compound sets on a regular base. If larger sets need to be analysed, feel free to contact our assay experts.

The concentration is mostly between 0.1 and 2 mg/ml but can be adjusted according to your needs. The concentration will be determined by either BCA, adsorption at 280nm or densitometric determination after Coomassie staining.

Yes, in most cases we use cleavable tag sequences. But cleavage efficiency can be influenced by the target protein and has to be tested.

We use many different tags according to your needs. Very common tags for purification are the following: His, MBP, GST and Strep. But we use also different epitope tags or tags increasing the solubility or the localization of the target protein.

For the production of glycosylated proteins we offer expression in mammalian cell lines, Yeast and in Baculo system (insect cells).

Depending on the expression system and the purification process, the timeline for generating a protein is starting at 20 business days.

No, generally we do not add preservatives to the proteins since they may influence the protein activity. But if you have a preferred storage buffer, we can use your buffer.

Yes, generally expression levels are higher after codon optimization. Therefore, we offer codon and DNA structure optimization within a project.

Our standard priority shipment will be on dry ice. If required, we offer shipment on blue ice, at -20°C or even at room temperature (lyophilized proteins only) as well.

Generally, we offer native purification conditions. Depending on target protein properties, denaturing purification can also be offered for proteins.

We can additionally offer sterile filtration if you require this service.

Posttranslational modifications like glycosylation or very rigid protein structures may have influences on protein properties and running behaviour. Sometimes proteins run higher, seldom lower and sometimes they run as a smear.

During initial free individual consulting, our scientific experts run a feasibility check (e.g. literature check, database check, product check, etc.) with your protein sequence to define the ideal service strategy for your specific target. The results will be discussed and you will receive a quote for the process proposed.

This will be decided by an initial feasibility check free of charge. Most important, the protein should be soluble and not a full length membrane protein. But the soluble domain of a membrane protein should work, for example.

We only need the amino acid sequence or uniprot reference number of your desired protein.

We have successfully expressed all protein classes in different expression systems and different expression strategies.

Yes, we offer molecular biology services like gene synthesis as addition to protein production services.

You will receive the purified protein including a datasheet. For more complex projects, you will also receive an additional and detailed report.

Our minimum guaranteed purity is > 75% and in many cases we have > 95% by only one purficiation step. The final purity can be further increased by an additional purification step.

We guarantee the following minimal quantities: 3 mg for proteins produced by E.coli, 1 mg for labelled proteins produced by E.coli and 1 mg for proteins produced within a mammalian package. If a pilot study is included, guaranteed delivered amount can be extended.

If you need both, your recombinant protein and a corresponding antibody, our guaranteed antibody package will be the best service choice for you.

For generation of glycosylated proteins we offer different expression systems like Yeasts, insect cells (Baculo) or also mammalian cell lines. Depending on the nature of your target protein and your exact needs, we will propose the most suitable expression host and system for your project.

A fixed price package for generation of a monoclonal antibody does not exist, since these types of services are individually designed in collaboration with our partner for immunization according to your exact needs.

Yes, we also guarantee the quality of an antiserum generated (polyclonal antibody). The final titer will be tested by ELISA.

Double and triple labeling are possible. We offer the following labels, which can be combined in different combinations: 15N, 13C, D2O.

Yes, we offer various recombinant proteins off the shelf. Have a look at our continuously growing catalog.

The typical yield is not easy to predict, but can reach up to gram-quantities per liter of high-cell density fermentation runs (E.coli and Yeast). The scale can reach up to 8 l for bioreactors and 10 l for cell culture.