title: Current methods for the synthesis of homogeneous antibody-drug conjugates.
abstract: Development of efficient and safe cancer therapy is one of the major challenges of the modern medicine. Over the last few years antibody-drug conjugates (ADCs) have become a powerful tool in cancer treatment with two of them, Adcetris® (brentuximab vedotin) and Kadcyla® (ado-trastuzumab emtansine), having recently been approved by the Food and Drug Administration (FDA). Essentially, an ADC is a bioconjugate that comprises a monoclonal antibody that specifically binds tumor surface antigen and a highly potent drug, which is attached to the antibody via either cleavable or stable linker. This approach ensures specificity and efficacy in fighting cancer cells, while healthy tissues remain largely unaffected. Conventional ADCs, that employ cysteine or lysine residues as conjugation sites, are highly heterogeneous. This means that the species contain various populations of the ADCs with different drug-to-antibody ratios (DARs) and different drug load distributions. DAR and drug-load distribution are essential parameters of ADCs as they determine their stability and efficacy. Therefore, various drug-loaded forms of ADCs (usually from zero to eight conjugated molecules per antibody) may have distinct pharmacokinetics (PK) in vivo and may differ in clinical performance. Recently, a significant progress has been made in the field of site-specific conjugation which resulted in a number of strategies for synthesis of the homogeneous ADCs. This review describes newly-developed methods that ensure homogeneity of the ADCs including use of engineered reactive cysteine residues (THIOMAB), unnatural amino acids, aldehyde tags, enzymatic transglutaminase- and glycotransferase-based approaches and novel chemical methods. Furthermore, we briefly discuss the limitation of these methods emphasizing the need for further improvement in the ADC design and development.
authors.affiliation: Faculty of Biotechnology, Department of Protein Engineering, University of Wroclaw, Joliot-Curie 14a, 50-383 Wroclaw, Poland.
authors.forename: Alicja M
authors.initials: AM
authors.lastname: Sochaj
authors.affiliation: Faculty of Biotechnology, Department of Protein Engineering, University of Wroclaw, Joliot-Curie 14a, 50-383 Wroclaw, Poland.
authors.forename: Karolina W
authors.initials: KW
authors.lastname: Świderska
authors.affiliation: Faculty of Biotechnology, Department of Protein Engineering, University of Wroclaw, Joliot-Curie 14a, 50-383 Wroclaw, Poland. Electronic address: otlewski@biotech.uni.wroc.pl.
authors.forename: Jacek
authors.initials: J
authors.lastname: Otlewski
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D000970:Antineoplastic Agents; D001709:Biotechnology / Q000379:methods*; D016503:Drug Delivery Systems; D006801:Humans; D018796:Immunoconjugates; D051379:Mice; D009369:Neoplasms / Q000188:drug therapy; D015202:Protein Engineering / Q000379:methods*; D051381:Rats
chemical_list: D000911:Antibodies, Monoclonal; D000970:Antineoplastic Agents; D018796:Immunoconjugates
keywords: ADCs; Cytotoxic agents; Homogeneity; Monoclonal antibodies; Site-specific conjugation; Targeted cancer therapy