Synthesis of Heterobifunctional Protein Fusions Using Copper‐Free Click Chemistry and the Aldehyde Tag

• DOI: 10.1002/anie.201108130
• Publication Date: 2012-03-12T07:53:29Z
• Authors: Jason E. Hudak; Robyn M. Barfield; Gregory W. de Hart; Patricia Grob; Eva Nogales; Carolyn R. Bertozzi; David Rabuka
• Keywords: Models, Molecular; Aldehydes; 01 natural sciences; Communications; Antibodies; Maltose-Binding Proteins; Protein Structure, Tertiary; 0104 chemical sciences; 3. Good health; Microscopy, Electron, Transmission; Growth Hormone; Molecular Probes; Humans; Click Chemistry; Copper

Heterobifunctional protein fusions are gaining interest as next-generation biopharmaceuticals.1–5 Combining proteins with disparate functions can enable multidrug therapy with a single chemical entity,6, 7 add a targeting element to an otherwise nonspecific therapeutic,8, 9 or improve the pharmacokinetic profile of a rapidly cleared molecule.10, 11 Indeed, heterobifunctional proteins, such as immunoglobulin G (IgG) Fc domain fusions, are among the top-selling biotherapeutics on the market today.12 These biomolecules are primarily generated as genetic fusions. The DNA sequences that encode the individual protein components are fused in tandem to direct the expression of a single polypeptide that comprises the two proteins joined together at their N and C termini, respectively. However, this limited topology is not ideal for every protein combination, as some polypeptides require unmodified termini for optimal bioactivity13 or can suffer from expression difficulties as a result of folding and processing issues.3, 14, 15