The advantages and disadvantages of bioorthogonal proteins

Groenewold, G.J.M. (2021)

Doctoral Thesis

This thesis describes the use of bioorthogonal proteins in immunological settings. It provides an introduction towards the field of protein modification, which was used throughout the thesis for the expression of proteins containing unnatural amino acids. It challenges this protein expression and the subsequent purification. With this successfully challenged, the use of the protein was assessed in antigen activation studies. It was analyzed whether the unnatural amino acids were tolerated by the T cell receptor and the ligation handles of the unnatural amino acids were subsequently used for visualization purposes. Furthermore, this thesis describes the possible use of azido HRP as a new tool for immunohistology.

• Unnatural amino acid
• Protein expression
• Protein purification
• Click chemistry
• Bioorthogonal ligation reactions
• Chemical immunology

• All authors: Groenewold, G.J.M.
• Supervisor: Kasteren, S.I. van; Overkleeft, H.S.
• Committee: Aerts, J.M.F.G.; Stelt, M. van der; Boyle, A.L.; Wezel, G.P. van; Bogaart, G. van den; Scheffer, D.J.
• Qualification: Doctor (dr.)
• Awarding Institution: Leiden Institute of Chemistry (LIC), Faculty of Science, Leiden University
• Date: 2021-02-17
• ISBN (print): 9789464212143