Innovative bioanalytical approaches to support the development of New Biological Entities (Protein, Antibodies, ADC (?)

The scope of this presentation is to give an overview of the different bioanalytical approaches used to support New Biological Entities (NBE) and Antibody Drug Conjugates (ADC) projects. Ligand Binding Assays are considered the gold standard for the determination of the concentration of proteins and antibodies in pharmacokinetic (PK) and toxicokinetic (TK) studies (serum, plasma); different approaches will be presented including ELISA, Gyrolab, Mesoscale Discovery (MSD), and Simoa Quanterix technologies. Immunogenicity risk associated with the development of anti-drug antibodies (ADAs) following administration of biotherapeutics is considered one of the main issues in the development of New Biological Entities. For this reason, different approaches to detect ADA in in-vivo studies will be presented. LC-MS technique is also emerging as an alternative tool for the determination of the concentration of total Antibody in biological samples. In addition, LC-MS/MS techniques can be employed to support in vivo ADC pharmacokinetics studies: they can be dedicated to the quantitation of the conjugated toxin, detecting the active molecule, after the capture of the entire ADC and enzymatic cleavage of the linker. Moreover, LC-MS/MS methods are used to quantify the unconjugated toxin potentially released from the ADC. Cell-based assays are playing a crucial role in the development of NBEs. Different approaches based on flow cytometry will be presented for the understanding of the mechanism of action of novel drugs in in-vitro (target internalization) and in-vivo (receptor occupancy) studies. Finally, an overview of the pharmacokinetic properties of NBE will be provided including the different data analysis approaches used for PK parameters determinations


P. Balestra, E. Bertotti, I. De Salve, A. Paoletti, F. Riccardi Sirtori


medicinal chemistry