Simponi ADCC and CDC Assays

BioOutsource has the experience, expertize and capabilities to provide the complete ADCC and CDC testing solutions for Simponi using our off-the-shelf methods.  For the methods to be successful, Simponi CDC and ADCC assays require TNF-Alpha to be present as the membrane-bound form, at sufficiently high levels:  a cell line engineered to express membrane-bound TNF-Alpha is required for these assays. Our classical ADCC assays are performed using effector cells (PBMCs or enriched Natural Killer (NK) cells) from genotyped donors that are harvested on the day of assay.

The Simponi CDC and ADCC bioassays measure the combined activity of the Fab binding and effector functions of Simponi, mediated through an initial interaction with C1q (CDC) or the Fc-Gamma Receptor IIIa (FcRIIIa) (ADCC). BioOutsource is industry experts in the development, optimization and qualification/validation of bioassays and has applied this expertise to golimumab to offer ADCC and CDC assays to our customers. We have provided services to support clone selection, characterization and biosimilarity activities with assays performed using our existing methodology, adapted to incorporate the customer’s reference standard of choice. Where required, rapid assay qualification can be achieved using customer’s material.

The principle of both Simponi (golimumab) ADCC and CDC assays is that the target cell line, expressing the membrane-bound form of TNF-Alpha, is prepared in cell plates. A dilution series of Simponi is then prepared, added to the plate and allowed to bind to the target cells for a period of time. PBMCs (ADCC bioassay) or complement (CDC bioassay) are then added; this will bind to Simponi/membrane-bound TNF-Alpha complex to initiate a cell death cascade resulting in target cell lysis. The assays are performed with appropriate assay controls, utilizing a non-radioactive assay endpoint. Sample results can be reported as a percent of total cell death or as a relative potency measurement against the reference standard.

ADCC assays are well known for day to day variability arising from the donor PBMCs that are used in the methodology and BioOutsource have implemented a number of different strategies to control this variation. These include donor screening programmes so that assays are only performed using qualified individuals, and a standardized cell culture regime.

Contacts our experts to discuss your ADCC and CDC assay requirements for Simponi biosimilars.