Peptide facility

 

Services

We routinely use Fmoc-solid phase chemistry and have 2 Syro II robot synthesizers and a Intavis synthesizer equipped with real time Fmoc-monitoring. On one of the Syro's 48 peptides can be synthesized simultaneously on a typical 25 µmol scale. The other Syro can synthesize 6 X 96 peptides in up to 4 µmol amounts in one run for screening purposes. The Intavis synthesizer is used for synthesizing larger amounts of peptides and for fine-tuning difficult sequences using the Fmoc-monitoring methodology.

 Besides "straight forward" peptides it is also possible to synthesize specifically modified peptides.

• Ubiquitin(like) and derivatives thereof
• Variations at the N-terminus; biotinylation (w/wo a spacer), acetylation, fluorophores etc.Phosphorylated peptides
• Peptides with a side chain modification (such as thiolysine e.g.)
• UV cleavable peptides
• Unnatural amino acid incorporation
• Alanine scans

Equipment

Being both an organic chemistry lab and a biochemistry/chemical biology lab working with reagents ranging from in house prepared small molecule inhibitor libraries all the way up to activity based probes consisting of modified di-ubiquitin proteins, we routinely use a wide variety of equipment:

Peptide synthesizers

• MultiSynTech Syro II - 1 x 24 parallel synthesizer (each 40 umol)
• MultiSynTech Syro II - 6 x 96 parallel synthesizer (each 4 umol)
• Intavis MultiPep CF 

Liquid Chromatography - Mass Spectrometers (LC-MS)

• WATERS Acquity H-class UPLC coupled to a LCT-premier Mass Spectrometer
• WATERS Acquity H-class UPLC coupled to a high resolution XEVO G2S-XTOF Mass Spectrometer 

HPLC-systems

• WATERS 2545 LCMS coupled to a 3100 Mass Spectrometer; mass triggered collection during HPLC purification mostly used for small molecules.
• WATERS 2535 LCMS coupled to a 2545 UV-VIS detector; routinely used for large scale purifications of Ubl's 
• WATERS Acquity H-class UPLC; routinely used for peptide analysis
• Shimadzu LC-20 LCMS system; routinely used for small scale purifications of peptides and Ubl's

FPLC-systems

• 2x BUCHI-C620 automated colomn set-ups for straight phase silica separations
• BIORAD NGC system for cat-/anion exchange and size exclusion gelfiltration

NMR-system

• Bruker Ultrashield 300 MHz

 

CONTACT

Cami Talavera Ormeno
E-mail: peptides@lumc.nl
Tel: +31 (0) 71 526 8730

 

          

Cami Talavera Ormeno                    

Peptide Facility
Department of Cell and Chemical Biology (CCB)
Leiden University Medical Center (LUMC)