| Autor: |
Jan, Grünewald, Ansgar, Brock, Bernhard H, Geierstanger |
| Rok vydání: |
2019 |
| Předmět: |
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| Zdroj: |
Methods in molecular biology (Clifton, N.J.). 2012 |
| ISSN: |
1940-6029 |
| Popis: |
4'-Phosphopantetheinyl transferases (PPTases) have been employed by researchers as versatile biocatalysts for the site-specific modification of numerous protein targets with structurally diverse molecules. Here we describe the use of these enzymes for the production of homogeneous antibody-drug conjugates (ADCs), which have garnered much attention as innovative anticancer drugs. The exceptionally broad substrate tolerance of PPTases allows for one-step and two-step conjugation strategies for site-specific ADC synthesis. While one-step conjugation involves direct coupling of a drug molecule to an antibody, two-step conjugation provides increased flexibility and efficiency of the conjugation process by first attaching a bioorthogonal chemical handle that is then used for drug molecule attachment in a second step. The aim of this chapter is to outline detailed protocols for both labeling procedures, as well as to provide guidance on enzyme and substrate preparation. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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