Welcome to ORGC, where we take pride in offering tailer-made solutions for for Nuclear Magnetic Resonance (NMR), Mass Spectrometry (MS), and Peptide Design applications. For inquiries, feel free to reach out to Prof. Dr. Steven Ballet at steven.ballet@vub.be. We look forward to assisting you in your scientific endeavors.
NMR Recording & Analysis
NMR data are recorded on a Bruker AscendTM 400 magnet equipped with an Avance NEO console, a 5 mm High-Resolution BroadBand Observe (HR-BBO) iProbe operating at 400.20 MHz (1H), and a fully automated SampleCase sample changer able to accommodate 24 samples. Spectra are routinely recorded at 300 K using standard Bruker pulse programs, making use of Topspin 4.5 in an ICON-NMR environment. Experiments involving 1H, 2H, 13C, 15N, 19F, 29Si and 31P can be performed on a routine basis, other nuclei can be considered on specific requests. A set of routinely available 1D and 2D spectra include the following:
- 1D 1H (the delay between scans can be increased (standard 1 s) depending on sample necessities (e.g. polymers, quantitative 1H))
- 1D 13C APT (outcome similar to a normal 13C and DEPT, including quaternary carbons)
- 1D 13C with 1H decoupling
- 1D 13C DEPT
- 1D 19F with/without 1H decoupling
- 1D 31P
- 2D 1H-1H COSY
- 2D 1H-1H TOCSY
- 2D 1H-1H NOESY or ROESY (ROESY for MW 1000-2000, NOESY for others; mixing time dependent on molecular weight)
- 2D 1H-13C or 1H-15N HSQC
- 2D 1H-13C HMBC
- 1D and 2D experiments involving water suppression.
Experiment time is prone to available sample material. Reasonable measurement time can be expected for amounts >5 mg (1H) and >20 mg (13C). Samples should be supplied in 5 mm NMR tubes (minimal specifications available upon request) containing ca. 40 mm of liquid sample. Sample preparation can be performed upon request, wherefore the following solvents are directly available (but not limited to): CDCl3, DMSO-d6, CD3CN, MeOD-d4, D2O, CD2Cl2. Additionally, spectra can be recorded in water upon addition of 10% D2O, requiring the use of water suppression methods (presaturation or excitation sculpting).
Additional experiments (e.g. 2D experiments considering other nuclei), use of specific parameters/modifications or measurement of non-routine nuclei can be considered for specific requests. Variable temperature measurements (-40 to 120°C) can be performed with heated/cooled pressurized air or nitrogen gas upon use of the internal probe heating system and/or a liquid nitrogen heat exchanger, depending on preferred temperatures.
Multiple options are available, considering sample preparation, experiment choice, parameter choice and/or data analysis (including full assignments).
Mass Spectrometry Recording & Analysis
The Synapt XS system (Waters Corporation) is a highly advanced mass spectrometry system equipped with a quadrupole and an extended Time-of-Flight (ToF). The system is compatible with an Electrospray Ionization (ESI), Desorption Electrospray Ionization (DESI) and Matrix-Assisted Laser Desorption/Ionization (MALDI) sources. Besides routine LC/MS, (HR)MS and MS/MS measurements, the Synapt XS is also compatible with mass spectrometry imaging applications (MSI). MSI is a promising, relatively new imaging technique with the great advantage that it does not require the presence of a label (e.g. radio-isotope, fluorophore) to visualize the compound of interest, hereby preventing potential non-specific binding or changes in target binding and functionality. Possible MSI applications include biodistribution studies, compound localization studies and biomarker identification.
Key applications:
- ESI mode: Analysis of small molecules and peptides (MW < 2-3 kDa)
- LC/MS
- LC/HRMS
- MS/MS
- DESI mode: Analysis of metabolites, lipids and peptides (MW < 2-3 kDa)
- MS
- MS/MS
- Mass spectrometry imaging applications (tissue sections; no whole-body imaging)
- MALDI mode: Analysis of lipids, peptides and proteins (MW up to ca. 7 kDa, option to increase if required)
- MS
- MS/MS
- Mass spectrometry imaging applications (tissue sections; no whole-body imaging)
Peptide Design & Synthesis
The Ballet lab enjoys a prominent position among peptide-chemistry research groups within Belgium and Europe and, as such, is well-suited to provide high-quality peptide design and synthesis. The group benefits from possibilities for both manual and automated synthesis. When required, a mixture of the two is also possible. For instance, the automated synthesis of a peptide with manual cyclizations, late-stage modifications or the coupling of non-canonical amino acids, chelators, tags etc. For the latter, the group is well-versed in the design and synthesis of such non-canonical amino acids, particularly those that introduce a conformational constraint into the primary sequence. The group is capable of full purification by preparative-HPLC as well as the full characterization of the synthesized sequences. In addition, structure determination is also possible for peptides with unknown identity (please see our NMR Services list for more information).
Non-exhaustive list of services:
- (Automated) synthesis of peptides up to ca. 50 AA*
- Cyclizations (various strategies depending on sequence identity)
- Incorporation of non-canonical amino acids
- Incorporation of visualisation tags, chelators
- Synthesis of non-canonical amino acids
- Peptide purification and lyophilization
- Structural characterization
- Peptide structure determination
- Consultancy services on peptide design and synthesis
* for longer lengths, we are open to discuss alternative synthetic methods to achieve this.