TSKgel® FcR-IIIA-NPR is based on a recombinant Fc gamma IIIa receptor ligand immobilised on a non-porous polymer particle. It allows fast assessment of biologic activity of monoclonal antibodies.
- Easy and reproducible HPLC analysis based on FcγRIIIa affinity of mAbs
- Unique glycoprotein elution profile of IgG allows assessment of ADCC activity
- Applicable to purified samples and cell culture supernatant alike
- Fast cell line screening, upstream development, lot-to-lot comparison
Fcγ IIIa receptor plays a key role in antibody-dependent cellmediated cytotoxicity (ADCC). ADCC is a crucial mechanism of action (MoA) of anti-tumor therapeutic antibodies. The Fc-glycans of antibodies are known to play an important role in Fc-mediated effector functions. Hence, separation patterns of therapeutic antibodies on TSKgel® FcR-IIIA-NPR can be correlated to Fc N-glycans. Terminal galactose residues increase affinity to FcR gamma while core fucose residues reduce it. This correlates with the known influence of galactose and fucose on ADCC activity. Accordingly, early eluting peaks of TSKgel® FcR-IIIA-NPR represent glycoforms with low ADCC activity while late eluting peaks represent glycoforms with high ADCC activity.
A rapid thirty-minute separation allows the analysis of large numbers of mAb samples to gain valuable first information on the distribution of glycoforms and expected ADCC activity. This fast and efficient method can be applied to purified samples and supernatant alike and can therefore be used in many phases of development and production such as cell line screening in early R&D, biosimilar/originator comparison, upstream development and optimisation, monitoring of glycoengineering, or lot-to-lot comparison in QC.
The stationary phase utilises a non-glycosylated, recombinant, human FcγRIIIa protein bound to a non-porous polymer bead and is packed in PEEK hardware. This ligand was engineered to have inherently high stability and nearly identical selectivity to wild type FcγIIIa receptor.
