A presentation at the Immunologisk Selskab meeting in 2006 about Tapasin and MHC-I antigen presentation

2. Advertisement: My PhD project Gustav Røder Panum Institute (by Søren Buus) University of Copenhagen Tapasin mediated receptor editing and quality control of major histocompatibility complex class I molecules CONTACT ME: [email_address] OR http://www.tapasin.com

4. The Human MHC Project Cloning Expression Purification Active HCs MHC-I Heavy Chain Manufacturing

5. Production of recombinant HLA complexes

6. Production of recombinant HLA complexes

7. Production of recombinant HLA complexes Jumping from one MHC-I allele to another by introducing mutations: These constructs are then ‘ codon optimized’ for efficient Expression in E.coli

8. Production of recombinant HLA complexes Purification by IMAC

9. Production of recombinant HLA complexes Ferre et al . Protein Sci. 2003, 12:551 Purification by HIC Fractions tested in RIA or ELISA

10. Production of recombinant HLA complexes Denatured MHC Heavy Chain Folded ß2-microglobulin Peptide Mixing Pot Size Exclusion Chromatography Concentration > 3 mg/mL

11. Crystallization and structure determination Hanging Drops MHC-I Complex

12. Crystallization and structure determination Model Building

13. Crystallization and structure determination

14. Crystallization and structure determination

15. Crystallization and structure determination ILGPPGSVY LEKARGSTY Epstein-Barr virus nuclear protein 3 EBNA3A (406-414) Human ubiquitin Conjugating enz. (83-91)

16. Modeling and structure based affinity prediction

17. Acknowledgements IMMI: Lotte Nielsen, Sune Justesen, Kasper Lamberth, Søren Buus DFH: Thomas Blicher, Ole Kristensen, Jette Kastrup, Michael Gajhede