Peptide facility

Peptides are routinely synthesized in the range of 5-80 amino acids, in an amount of 1-200 µmol (smaller and larger amounts are also possible). All peptides are analyzed by LC-MS and if needed purified by RP-HPLC. For analytical purposes the peptide core facility has a Waters liquid chromatography electro spray mass spectrometer at its disposal. More than 6000 peptides are synthesized on a yearly basis.

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 

Mass Spectrometers

  • WATERS Alliance  LCMS 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 1525 EF LCMS coupled to a 2487 dual labda Absorbance Detector; routinely used for peptide purifications
  • 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 Paul Hekking

Cami Talavera Ormeno    Paul Hekking                       Dris El Atmioui


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

Groups

Collaborate with us

Looking for information on one of our topics, a new place to conduct your research or experienced research to join forces with?  Feel free to contact us.!

Read more