HPLC is the workhorse for isolation and analysis. Instruments in use include Shimadzu LC10 and LC20 systems equipped with autosamplers and uv/diode array detectors, a Waters 600 preparative system (flow rates <20 ml/min). We use columns ranging from 20 mm inner diameter down to 0.075 mm, that is we separate and analyse peptides from µmole to femtomole scale!
Two mass spectrometers are currently in use. The ThermoFinnigan LCQ Deca ion trap is used as a walk up instrument. Nanospray and HPLC can be used as inlets, and the instrument is capable of multistage MS/MS experiments.
The second instrument is a Waters Micromass QTof micro. A nanoAcquity UPLC is used as the standard inlet, and a lockmass source helps with accuracy. Highly reproducible separations and accurate mass measurements (<5 ppm) can thus be achieved using a minimum amount of analyte.
We use a CEM Liberty1 automatic peptide synthesizer and manual peptide syntesis. Fmoc-based chemistry is used for synthesis of small and larger (35-40 mer) peptides. Circular peptides are made using peptide thiosters combined with native chemical ligation, or by enzymatic ligation.
Biological and biophysical assays
We have expertise and inhouse facilities for tests of antibacterial and antifungal activities, and biophysical membrane studies. Instruments include a Hitachi F-2700 fluorescence spectrophotometer, a Thermo Nanodrop spectrophotometer. Lately, we installed a Direct Detect FT-IR system for accurate determinations of peptide and protein concentration.
We welcome inquiries about collaborative projects; and we can offer limited services for peptide synthesis. Contact Ulf Göransson for more information.