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Locked by Design: A Conformationally Constrained Transglutaminase Tag Enables Efficient Site‐Specific Conjugation |
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| Authors: | Dr. Vanessa Siegmund Dr. Stefan Schmelz Stephan Dickgiesser Jan Beck Aileen Ebenig Heiko Fittler Dr. Holm Frauendorf Dr. Birgit Piater Dr. Ulrich A. K. Betz Dr. Olga Avrutina Dr. Andrea Scrima Prof. Dr. Hans‐Lothar Fuchsbauer Prof. Dr. Harald Kolmar |
| Affiliation: | 1. Clemens‐Sch?pf‐Institut für Organische Chemie und Biochemie, Technische Universit?t Darmstadt, Alarich‐Weiss‐Stra?e 4, 64287 Darmstadt (Germany);2. Arbeitsgruppe Strukturbiologie der Autophagie, Abteilung Struktur und Funktion der Proteine, Helmholtz‐Zentrum für Infektionsforschung, Inhoffenstr. 7, 38124 Braunschweig (Germany);3. Institut für Organische und Biomolekulare Chemie, Zentrale Analytik/Massenspektrometrie, Georg‐August‐Universit?t G?ttingen, Tammannstr. 2, 37077 G?ttingen (Germany);4. Merck KGaA, Frankfurterstr. 250, 64293 Darmstadt (Germany);5. Fachbereich Chemie‐ und Biotechnologie, Hochschule Darmstadt, Schnittspahnstra?e 12, 64287 Darmstadt (Germany) |
| Abstract: | Based on the crystal structure of a natural protein substrate for microbial transglutaminase, an enzyme that catalyzes protein crosslinking, a recognition motif for site‐specific conjugation was rationally designed. Conformationally locked by an intramolecular disulfide bond, this structural mimic of a native conjugation site ensured efficient conjugation of a reporter cargo to the therapeutic monoclonal antibody cetuximab without erosion of its binding properties. |
| Keywords: | bioconjugation transglutaminase antibodies protein engineering site‐specific ligation |