Peptide production group

Our peptide group routinely produces peptides for several research groups within the DKFZ (for internal use only).

Validation of immune epitopes relies on fast and cost-effective production of short (8 - 11 aa for MHC I epitopes) and long (13 - 25 aa for MHC II epitopes) peptides. The peptide group has set up a workflow for parallel synthesis and purification of hundreds of peptides each year. Theses peptides are applied in research projects from various DKFZ groups for multiple purposes such as MHC-multimer staining, peptide-pulsed antigen-presenting cells (APCs) for functional T cell assays as well as murine peptide vaccination. Customized peptides (e.g. fluorophore-labelled) are also produced on request.

For solid phase synthesis Fmoc chemistry is used in a fully automated multiple synthesizer. Synthesis is carried out on preloaded Wang-resins.

Purification is done by preparative HPLC on a Kromasil 100–10C 10 µm 120A reverse phase column using an eluent of 0.1% trifluoroacetic acid in water (A) and 80% acetonitrile in water (B). The peptide is eluted with a successive linear gradient of 25% B to 80% B. Fractions corresponding to the purified peptide are lyophilized and characterized with analytical HPLC and MS.

Peptide-vaccination with patient-individual cancer neoepitopes is a promising approach for tumor treatment and can induce long-lasting immune responses in the patient. For vaccination strategies peptides have to be produced under GMP conditions with strict hygienic standards including a cleanroom environment and extended QC measure-ments. The peptide group currently adapts the peptide production process to fulfill GMP requirements and synthesize peptides as an active substance for clinical trials.