Skip Navigation | ANU Home | JCSMR | CMHS | Search ANU

Tetramer synthesis, Antibody purification, Protein Expression and Purification

The BRF has recently established a new MHC-Tetramer synthesis service. We are able to provide custom synthesis of soluble MHC-peptide tetramer reagents to researchers at the ANU. Tetramers and/or their derivatives are provided for research purposes only. Their use for commercial purposes is prohibited.

Although synthesis of Tetramers will be the main service offered by this lab, facilities and expertise are also available for antibody purification, and general protein experession and purification.

Services offered:

• MHC-tetramer synthesis (full service)
• Antibody purification (limited service)
• Protein expression and purification (training provided to assist you in using our facilities)
• Advice on all aspects of protein expression, purification and chromatography (Peter Milburn)

Tetramer synthesis

Currently available MHC Class I Alleles.

Murine

H-2D(b)

H-2D(d)

H-2K(b)

H-2K(d)

H-2L(d)

At present reagents are labeled with streptavidin-allophycocyanin (SA-APC).

Quality control: All tetramers are purified by size exclusion and ion exchange chromatography and each tetramer is tested for level of biotinylation.

Synthesis: The amount of biotinylated monomer that is obtained is dependent on a number of factors. One factor that determines the success of tetramer production is the binding affinity of the peptide for the MHC-class I molecule. The yield of monomer will therefore be variable for each folding and will determine the number of aliquots available for subsequent tetramerizations. Biotinylated monomer will be aliquoted and stored in 50 ug units.

Pricing: This is based on the cost of the initial folding and the scale of tetramerization/label required. Subsequent tetramerizations of aliquots of monomer from the same folding are available at a reduced price, provided no new monomer preparation is required. These prices are based on our current costs to produce tetramers.

For example - if the folding yields 1000 ug ( 5 x 200 ug aliquots), the cost for the first prep will be the cost to produce biotinylated monomer plus labelling at the scale of your choice (currently 50, 100, 200 ug). Choice of label will also affect the cost. For subsequent tetramerizations from stored monomer you will only pay for the labelling/tetramerization.

Peptides: Can be ordered through the BRF's Peptide Synthesis service. Peptides should be HPLC purified and in powder form. 10 mg of each peptide are required for each tetramer production. Analytical HPLC and MS should be performed on all peptides prior to use.

---

PLEASE NOTE: All tetramers synthesised in the BRF must be acknowledged in the ”Materials and Methods” section of your paper and reference to use of the BirA enzyme in the synthesis MUST be cited (see web site for eg).
O’Callaghan, C.A., Byford, M.F., Wyer, J.R., Willcox, B.E., Jakobsen, B.K., McMichael, A.J. and Bell, J.I (1999). BirA Enzyme: Production and Application in the study of membrane receptor-ligand interactions by site-specific biotinylation. Anal. Biochem. 266, 9-15.

---

Ordering
Please complete a Tetramer Synthesis order form and discuss your requirements with Kerong BEFORE placing your first order. Ensure that your peptide is clearly labelled with your name and peptide ID. You will be informed by email when the tetramer is ready for pickup.

Pricing

Contact

Dr Kerong Zhang
Tel: 6125 9539
email: Kerong.Zhang@anu.edu.au