Mouse Anti-b96.11 Recombinant Antibody (clone 8E6G4)

Purified MAb 8E6G4, supernatant of hybridoma 8E6G4, or BSA were analyzed for binding by b96.11 by Dot blot. From left: supernatant of hybridoma 8E6G4, purified MAb 8E6G4, and BSA.

Wang, X., Zhang, A., Liu, Y., Chen, S., Feng, Z., Shang, W., ... & Hampe, C. S. (2012). Anti-idiotypic antibody specific to GAD65 autoantibody prevents type 1 diabetes in the NOD mouse. PloS one, 7(2), e32515.

Binding of MAb 8E6G4 and supernatant of hybridoma 8E6G4 to b96.11 was analyzed by immunoprecipitation followed by Western blot analysis. Binding of MAb 8E6G4 to control antibody hLF served as negative control. From left: supernatant of hybridoma 8E6G4 with b96.11, MAb 8E6G4 with b96.11, and MAb 8E6G4 with hLF.

Wang, X., Zhang, A., Liu, Y., Chen, S., Feng, Z., Shang, W., ... & Hampe, C. S. (2012). Anti-idiotypic antibody specific to GAD65 autoantibody prevents type 1 diabetes in the NOD mouse. PloS one, 7(2), e32515.

MAb 8E6G4 binds to b96.11 with high affinity. Dissociation constant was calculated by non-competing ELISA. Different concentrations of b96.11 were tested for binding by MAb 8E6G4. Kd of each concentration was calculated. The Kd was calculated as follows: Kd = 2(nKd′-Kd)/(n-1), where n = [b96.11]′/[b96.11]. The average Kd calculated was 480.4 pM.

Wang, X., Zhang, A., Liu, Y., Chen, S., Feng, Z., Shang, W., ... & Hampe, C. S. (2012). Anti-idiotypic antibody specific to GAD65 autoantibody prevents type 1 diabetes in the NOD mouse. PloS one, 7(2), e32515.

A: GAD65 was translated in vitro using rabbit reticulocyte lysate. Different amounts (0–40 µl, corresponding to 0–160 ng) of recombinant GAD65 were incubated with 1.4 µg/ml MAb 8E6G4 and subsequently analyzed for binding to b96.11 by ELISA. Rabbit reticulocyte lysate alone was used as a negative control (open squares). Binding in the absence of competitor was set as 100%. B: Binding of monoclonal GAD65Ab b96.11 (black squares), b78 (open squares), and N-GAD65mAb (open circles) to radiolabeled GAD65 in the presence of the indicated concentrations of MAb 8E6G4 was determined. Binding is reported at percent binding, binding of the respective antibody to radiolabeled GAD65 in the absence of MAb 8E6G4 is set as 100%.

Wang, X., Zhang, A., Liu, Y., Chen, S., Feng, Z., Shang, W., ... & Hampe, C. S. (2012). Anti-idiotypic antibody specific to GAD65 autoantibody prevents type 1 diabetes in the NOD mouse. PloS one, 7(2), e32515.

NOD mice were injected with 50 and 100 µg MAb 8E6G4, presented as black circles and squares, respectively. Control animals were injected with PBS (white circles) or control mouse monoclonal antibody (white squares). Animals were weekly monitored for diabetes development. Cumulative incidence rate of diabetes development is presented as percentage.

Wang, X., Zhang, A., Liu, Y., Chen, S., Feng, Z., Shang, W., ... & Hampe, C. S. (2012). Anti-idiotypic antibody specific to GAD65 autoantibody prevents type 1 diabetes in the NOD mouse. PloS one, 7(2), e32515.

Representative images of pancreatic islets from animals injected with 50 µg (A) and 100 µg (B) MAb 8E6G4 IgG, or control animals (C). Pancreatic tissues were sectioned and stained with hematoxylin and eosin.

Wang, X., Zhang, A., Liu, Y., Chen, S., Feng, Z., Shang, W., ... & Hampe, C. S. (2012). Anti-idiotypic antibody specific to GAD65 autoantibody prevents type 1 diabetes in the NOD mouse. PloS one, 7(2), e32515.