Biointron has developed our own CHOK1-Fut8KO expression platform for afucosylated antibodies. Combined with our high-throughput recombinant antibody expression platform and cell culture technology, afucosylated antibodies can be rapidly produced for your research and industrial needs. Afucosylation has been shown to significantly boost antibody-dependent cell-mediated cytotoxicity (ADCC), opening up a realm of possibilities for targeted therapies by increasing effectiveness.
1: Glycosylation Profiling
This case study analyses glycosylation profiles by mass spectrometry on two types of cells. Fucose was not detected on modified cells in the N-glycan LC-MS – indicating that the CHOK1BN-Fut8KO cell can successfully express afucosylated antibodies. Here, C2193xxxx-1 is the standard CHOK1BN, C2193xxxx-2 is the CHOK1BN-Fut8KO.
Figure 1. C2193xxxx-1 is the standard CHOK1BN, C2193xxxx-2 is the CHOK1BN-Fut8KO.
2: Purity Verification of the Antibody Expressed by CHOK1BN-Fut8KO
High purity for antibodies expressed by CHOK1BN and CHOK1BN-Fut8KO. SDS-PAGE and SEC-HPLC verified that both antibody aggregation and purity met the >95% standard.
Comparing the ADCC effect of antibodies (Rituximab) expressed by the afucosylated host cell CHOK1BN-Fut8KO and antibodies expressed by CHOK1BN (as a control) on target cells through ADCC testing, the antibodies expressed by CHOK1BN-Fut8KO have significantly increased NK cell-mediated killing of target cells compared to antibodies from CHOK1BN. In addition, ADCC EC50 values were significantly improved.
4: Antibody Affinity Measuring
Using Biacore to measure the affinity between the antibody and the antigen FcγRIIIa, the afucosylated antibody expressed by CHOK1BN-Fut8KO can significantly increase its affinity for binding to the receptor, thereby enhancing the ADCC effect.
