Laboratory of New Methodologies of Peptide Synthesis
Department of Inorganic Chemistry
Faculty of Pharmacy
ABOUT US
The Laboratory provides services aimed at researchers and commercial customers. We perform contract-based syntheses, develop new methods and also optimize processes for peptide syntheses on solid media. We accept orders for commissioned syntheses on scales from 2 mg to as much as 10 g. The Laboratory provides peptides with documentation concerning purity (HPLC analysis) and identity (MS spectrum). We obtain linear peptides, cyclic peptides modified with non-protein amino acid residues, fatty acid residues and markers (e.g. biotin, rhodamine, fluorescein). Peptides of different purity levels are available as needed (crude, >80%, >95%). In addition, within the framework of cooperation with the University of Gdansk, there are additional tests available for the obtained compounds, including analyses of: NMR, IR, MADLI-TOF MS, elemental analysis and circular dichroism spectrum.
Synthesis methodology
The Laboratory specializes in synthesis by chemical methods on a solid polymer carrier, based on Fmoc/tBu methodology. We perform syntheses manually at room temperature or at elevated temperatures, using heating lamps and a fully automatic microwave peptide synthesizer. The laboratory is equipped with one of the latest microwave synthesizers, and its use allows a significant reduction in synthesis time, as well as optimizing the reaction conditions (temperature, reagent ratio, reaction time, etc.). Furthermore, one of the significant advantages offered by microwave synthesizers is reduced reagent consumption, demonstrating both economic and environmental significance.
Purification and peptide analysis
We purify the compounds using chromatographic techniques (mainly preparative reversed-phase HPLC, RP-HPLC). The Laboratory is equipped with a preparative chromatograph, allowing compounds to be purified on the milligram to gram scale. The compounds are purified in a water-acetonitrile solvent system, with the addition of trifluoroacetic acid, and then lyophilized. We analyze the purity of the obtained compounds by techniques using analytical HPLC and HPLC coupled to a mass spectrometer.
Difficult sequences and modifications
We specialize in the synthesis of the so-called difficult sequences, containing highly aggregating fragments. The Laboratory personnel has experience in the synthesis of cyclic peptides (cyclization between the N- and C-terminus, disulfide bonds), containing isotope-labeled amino acid residues, D-amino acids and non-protein fragments. Some of the possible modifications include derivatives: biotinylated peptides, containing fluorescent markers (rhodamine, fluorescein), lipidated peptides (N-terminal,as well as in the side chain), glycosylation or quaternization.
Replacement and analysis of counter-ions
Peptides obtained by SPPS are most often found in ionized form, as trifluoroacetate salts. The Laboratory offers the possibility of synthesizing peptides with other counter-ions. On request, we provide peptides as acetate or chloride salts. In addition, we have developed procedures for replacing counter-ions in peptides based on lyophilization, evaporation, chromatographic techniques, or the use of ion-exchange resins. The exchange level and counter-ion content is verified by ion chromatography. The Laboratory is equipped with an analytical ion chromatograph – Dionex ICS-5000+.
Development of new methods for peptide synthesis
We provide services to modify classical methodologies towards dedicated syntheses. We optimize customers’ syntheses in terms of rational use of equipment and the amount of reagents utilized. We support correct equipment selection and implement processes at the customer’s site.
Training activities
We provide fee-based courses in peptide synthesis, purification and analysis. The courses are conducted individually, as well as in groups of 2-3 people. As a part of the course, the participant learns the process for obtaining peptides by chemical synthesis from the planning stage to the final product. The participant independently synthesizes the peptide using an automated microwave synthesizer or performs a manual synthesis, separates the peptide from the resin, purifies them using a preparative HPLC kit, and analyzes purity and identity using HPLC coupled to a mass spectrometer.
Laboratory’s original research
We are conducting research on the use of elevated temperature in both the synthesis, purification and analysis of peptides. This research is aimed at a more economical and environmentally friendly approach to the process of obtaining highly purified peptides. This is due to the fact that increased temperature leads to accelerated peptide synthesis process, and also allows to increase the efficiency of individual reactions. In addition, the elevated temperature affects the compound retention in the RP-HPLC system, allowing to optimize the purification method, thus leading to reduced acetonitrile consumption.
CONTACT
Laboratory of New Methodologies of Peptide Synthesis
Department of Inorganic Chemistry
Medical Uniersity of Gdańsk
Dębinki 7 Street
80-211 Gdańsk
Department of Inorganic Chemistry
Medical Uniersity of Gdańsk
Dębinki 7 Street
80-211 Gdańsk
Photo Paweł Sudara/MUG and personal archive