TSKgel FcR-IIIA columns simplify Fc receptor affinity studies of antibodies
TSKgel FcR-IIIA columns are based on a recombinant Fc gamma IIIa receptor which is immobilized as a ligand on polymer particles. They separate monoclonal antibodies according their affinity to the receptor and enable a fast analysis of biological activity as well as fractionation along with biological activity.
Affinity chromatography based on Fc gamma IIIa receptor correlates to glycosylation and ADCC
Fc gamma IIIa receptor plays a key role in antibody-dependent cell mediated cytotoxicity (ADCC) which is a crucial mechanism of action of anti-tumor 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 can be correlated to Fc N-glycans and the ADCC activity of immunoglobulins. Early eluting peaks of TSKgel FcR-IIIA columns represent glycoforms with low ADCC activity while late eluting peaks represent glycoforms with high ADCC activity.
TSKgel FcR-IIIA-NPR for fast Fc receptor affinity analysis
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 optimization, monitoring of glycoengineering, or lot-to-lot comparison in QC.
TSKgel FcR-IIIA-5PW for collection of antibodies with differing Fc receptor affinities
The semi-preparative column can be loaded with sufficient material to result in fractions containing enough protein to use for further analyses. The resulting fractions are antibodies of different Fc receptor affinities and can be characterized, for instance as to their glycosylation status, ADCC activity or kinetic binding properties. This helps to understand the basis of Fc receptor affinity, antibody structure and function. Furthermore, data generated with fractions of the FcR-IIIA-5PW column underpin data obtained with FcR-IIIA-NPR column and validate correlations to glycosylation or ADCC.
FcR-IIIA affinity chromatography separates mAb glycoforms according to their ADCC activity |
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Column durability - analysis of CHO supernatant |
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