Mouse Anti-CD28 Recombinant Antibody (clone ANC28.1/5D10); F(ab')₂ Fragment (CAT#: DrMAB-981)

Figure 1 The CD28 superagonist ANC28.1 induces polyclonal T cell activation in vitro and leads to ZAP70 independent phosphorylation of LAT and PLCc1.

5x10⁴ freshly prepared human T cells per well were seeded in 96-well plates and treated with the indicated combinations of antibodies or were incubated in medium as a control.

Waibler, Z., Sender, L. Y., Merten, C., Hartig, R., Kliche, S., Gunzer, M., ... & Schraven, B. (2008). Signaling signatures and functional properties of anti-human CD28 superagonistic antibodies. PloS one, 3(3).

Figure 2 Induction of sustained calcium flux by human CD28 superagonistic antibody ANC28.1.

1x10⁶ freshly prepared Indo-1 loaded human T cells were stimulated with either a 1:50 v/v dilution of ascites fluid of CD3 mAb MEM92 (orange triangle), 10 mg/ml conventional CD28 mAb (CD28.2; blue triangle) followed by crosslinking with 20 mg/ml polyclonal goat anti-mouse antiserum (black triangle) or 10 mg/ml ANC28.1 (red triangle) followed by crosslinking with 20 mg/ml polyclonal goat anti-mouse antiserum (black triangle).

Waibler, Z., Sender, L. Y., Merten, C., Hartig, R., Kliche, S., Gunzer, M., ... & Schraven, B. (2008). Signaling signatures and functional properties of anti-human CD28 superagonistic antibodies. PloS one, 3(3).

Figure 3 Sustained calcium flux, T cell proliferation, and T cell signaling upon TGN1412 treatment.

5x10⁴ freshly prepared human T-cells/96-well were stimulated with the indicated concentrations of ANC28.1 or TGN1412 or were left untreated as a control. For crosslinking of ANC28.1, a polyclonal goat anti-mouse antiserum was used and crosslinking of TGN1412 was performed.

Waibler, Z., Sender, L. Y., Merten, C., Hartig, R., Kliche, S., Gunzer, M., ... & Schraven, B. (2008). Signaling signatures and functional properties of anti-human CD28 superagonistic antibodies. PloS one, 3(3).

Figure 4 Only engagement of CD81 by HCV-E2 or neutralizing Abs synergizes with ligation of CD28.

Human naive T cells were cultured with recombinant HCV envelope proteins immobilized on plate-bound anti-E2 plus soluble anti-CD28 (clone ANC28.1/5D10).

Serra, A., Nuti, S., Tavarini, S., Sammicheli, C., Rosa, D., Saletti, G., ... & Wack, A. (2008). Coligation of the hepatitis C virus receptor CD81 with CD28 primes naive T lymphocytes to acquire type 2 effector function. The Journal of Immunology, 181(1), 174-185.

Figure 5 ANC28 has properties of 'mitogenic' CD28 antibody.

CFSE-labeled PBMC stimulated with CD3, PHA, ANC28 or 28.2 alone, or CD28 mAb in combination with CD3 were analyzed for activation (CD25 up-regulation) and expansion (CFSE dye dilution) by flow cytometry on day 6.

Singh, M., Basu, S., Camell, C., Couturier, J., Nudelman, R. J., Medina, M. A., ... & Lewis, D. E. (2008). Selective expansion of memory CD4+ T cells by mitogenic human CD28 generates inflammatory cytokines and regulatory T cells. European journal of immunology, 38(6), 1522-1532.

Figure 6 Selective expansion of CD4+CD45RO+ cells by ANC28.

CFSE-labeled purified CD4⁺, CD8⁺ cells stimulated with 28.2 or ANC28.

Singh, M., Basu, S., Camell, C., Couturier, J., Nudelman, R. J., Medina, M. A., ... & Lewis, D. E. (2008). Selective expansion of memory CD4+ T cells by mitogenic human CD28 generates inflammatory cytokines and regulatory T cells. European journal of immunology, 38(6), 1522-1532.