Site-Specific Antibody-Drug Conjugation Using Microbial Transglutaminase
| Autor: | Stephan, Dickgiesser, Lukas, Deweid, Roland, Kellner, Harald, Kolmar, Nicolas, Rasche |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Chromatography
Chromatography Reverse-Phase Immunoconjugates Transglutaminases Molecular Structure Amino Acid Motifs Antibodies Monoclonal Catalysis Substrate Specificity Structure-Activity Relationship Mutagenesis Site-Directed Humans Amino Acid Sequence Hydrophobic and Hydrophilic Interactions Chromatography High Pressure Liquid |
| Zdroj: | Methods in molecular biology (Clifton, N.J.). 2012 |
| ISSN: | 1940-6029 |
| Popis: | Antibody-drug conjugates (ADCs) are a relatively young class of cancer therapeutics that combine the superior selectivity of monoclonal antibodies (mAbs) with the high potency of cytotoxic agents. In the first generation of ADCs, the toxic payload is attached to the mAb via chemical conjugation to endogenous lysine or cysteine residues providing only limited control over site specificity and drug-to-antibody ratio (DAR). The resulting product is a heterogeneous population of different ADC species, each with individual characteristics concerning pharmacokinetics, toxicology, and efficacy. Such diverse ADC mixtures are not only difficult to develop but are potentially also accompanied by a suboptimal therapeutic window. To overcome these limitations, alternative conjugation technologies have been developed that allow the production of tailor-made homogeneous ADCs. Due to its high specificity and robust applicability, microbial transglutaminase (mTG), a protein-glutamine γ-glutamyltransferase isolated from Streptomyces mobaraensis, emerged as a versatile tool for ADC manufacturing. Herein, we report a protocol for the site-specific, mTG-mediated modification of antibodies that allows the production of homogeneous and defined ADCs. Moreover, analytical methods for ADC characterization are provided. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|