Mouse Anti-CD3E Recombinant Antibody (clone UCHT1); Fab Fragment (CAT#: DrMAB-989)

Figure 1 Concanavalin A expanded γδ T cells staining

Concanavalin A expanded γδ T cells were stained with UCHT1, Fab, Fabred, or left untreated, followed by an APC-labeled anti-mouse IgG antibody as a secondary reagent. Fluorescence intensities were quantified by flow cytometry (n > 3).

Juraske, C., Wipa, P., Morath, A., Hidalgo, J. V., Hartl, F. A., Raute, K., ... & Pongcharoen, S. (2018). Anti-CD3 Fab fragments enhance tumor killing by human γδ T cells independent of Nck recruitment to the γδ T cell antigen receptor. Frontiers in immunogy, 9, 1579.

Figure 2 Close proximity between the TCR and Nck was detected by in situ PLA.

Zoledronate-expanded γδ T cells were either left untreated (−) or treated with 5 µg/ml UCHT1 in the absence or presence of 10 nM AX-024 at 37°C for 5 min. After fixation and permeabilization, PLA was performed using the primary antibodies goat anti-CD3ε (M20) and rabbit anti-Nck1, and the corresponding secondary antibodies. Nuclei were stained with DAPI.

Juraske, C., Wipa, P., Morath, A., Hidalgo, J. V., Hartl, F. A., Raute, K., ... & Pongcharoen, S. (2018). Anti-CD3 Fab fragments enhance tumor killing by human γδ T cells independent of Nck recruitment to the γδ T cell antigen receptor. Frontiers in immunogy, 9, 1579.

Figure 3 UCHT1 increased tumor killing by γδ T cells.

51Cr-labeled Daudi and Raji cells were incubated with zoledronate-expanded human γδ T cells without (−) or with the addition of 5 µg/ml UCHT1 using an effector to target (T cell to tumor cell) ratio of 12.5:1.

Juraske, C., Wipa, P., Morath, A., Hidalgo, J. V., Hartl, F. A., Raute, K., ... & Pongcharoen, S. (2018). Anti-CD3 Fab fragments enhance tumor killing by human γδ T cells independent of Nck recruitment to the γδ T cell antigen receptor. Frontiers in immunology, 9, 1579.

Figure 4 Using its SH3.1(Nck) domain Nck binds to the UCHT1-stimulated γδ T cell antigen receptor (TCR).

Zoledronate-expanded γδ T cells were left untreated (−) or stimulated with 5 µg/ml UCHT1 at 37°C for 5 min

Juraske, C., Wipa, P., Morath, A., Hidalgo, J. V., Hartl, F. A., Raute, K., ... & Pongcharoen, S. (2018). Anti-CD3 Fab fragments enhance tumor killing by human γδ T cells independent of Nck recruitment to the γδ T cell antigen receptor. Frontiers in immunology, 9, 1579.

Figure 5 Preparation of UCHT1 Fab fragments.

The anti-CD3ε antibody UCHT1 was digested with papain using a kit from Thermo Fisher Scientific. The Fab fragments were purified with protein A coupled beads.

Juraske, C., Wipa, P., Morath, A., Hidalgo, J. V., Hartl, F. A., Raute, K., ... & Pongcharoen, S. (2018). Anti-CD3 Fab fragments enhance tumor killing by human γδ T cells independent of Nck recruitment to the γδ T cell antigen receptor. Frontiers in immunogy, 9, 1579.

Figure 6 Fab and Fabred fragments enhanced tumor killing by γδ T cells.

51Cr-labeled Daudi and Raji cells were incubated with zoledronate or with 5 µg/ml UCHT1, 3.33 µg/ml Fab, or 3.33 µg/ml Fabred (these concentrations were chosen to obtain equimolar amounts of the different reagents). The effector to target ratio was 12.5:1.

Juraske, C., Wipa, P., Morath, A., Hidalgo, J. V., Hartl, F. A., Raute, K., ... & Pongcharoen, S. (2018). Anti-CD3 Fab fragments enhance tumor killing by human γδ T cells independent of Nck recruitment to the γδ T cell antigen receptor. Frontiers in immunogy, 9, 1579.

Figure 1 Mouse Anti-CD3E Recombinant Antibody (clone UCHT1); Fab Fragment (DrMAB-989) in SDS-PAGE

SDS-PAGE analysis of DrMAB-989 in non-reduced (Lane 1) and reduced (Lane 2) conditions. Gel stained for 30 minutes with Coomassie Blue. As a result, different β-mercaptoethanol-reduced proteins (Heavy chain and Light chain) migrate as about 50 kDa and 25 kDa, respectively.

The purity of DrMAB-989 was greater than 95% as determined by SDS-PAGE.

Figure 2 Mouse Anti-CD3E Recombinant Antibody (clone UCHT1); Fab Fragment (DrMAB-989) in HPLC

The purity of DrMAB-989 was greater than 95% as determined by SEC-HPLC.