TSKgel® FcR-IIIA-NPR is based on a recombinant FcγIIIa receptor ligand immobilized on a non-porous polymer particle. It allows fast assessment of biologic activity of monoclonal antibodies.
- Ligand: Modified recombinant FcγRIIIA (E. coli expression system, non-glycosylated)
- Base material: Polymethacrylate
- Pore size (mean): Non-porous
- pH stability: pH 4 - 8 (short term), pH 5 - 7 (long term)
- Base matrix: Nonporous resin, 5 µm
- Sample mass: 5 to 50 µg of IgG (recommended)
- Recommended buffer system A: 50 mM/L citrate buffer, 150 mM/L NaCl, pH 6.5
- Recommended buffer system B: 50 mM/L citrate buffer, 150 mM/L NaCl, pH 4.5
TSKgel FcR-IIIA-NPR column is used for high performance affinity chromatography. This column is designed for the separation of mAb efficacy variants on the basis of affinity of the N-linked glycosylation in the Fc Region of IgG1 -Fc for the recombinant FcγRIIIa stationary phase. The ligand is bonded to nonporous polymethrylate beads, providing efficient and rapid separation of mAb glycoforms. The rugged nature of the column facilitates analysis both prior to or after purification.
FcγIIIa receptor plays a key role in antibody-dependent cell mediated cytotoxicity (ADCC), which is a crucial mechanism of action of anti-tumor therapeutic mAbs. 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 and the ADCC activity of immunoglobulins. Early eluting peaks of TSKgel FcR-IIIA-NPR represent glycoforms with low ADCC activity while late eluting peaks represent glycoforms with high ADCC activity.
The rapid thirty minute separation allows the analysis of large numbers of mAb samples to gain valuable initial 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 and D, biosimilar/originator comparison, upstream development and optimization, monitoring of glycoengineering, or lot-to-lot comparison in QC. NPR stands for Non-Porous Resin.
TSKgel FcR-IIIA-NPR can be utilized for the following applications:
Comparison between biosimilar/biobetter and innovator reference product
QC Analysis of lot-to-lot difference for mAb drug products
Monitoring fermentation stage of cell culture media
Screening the potential of cell lines for ADCC activity
