NeuroMab™ Anti-Amyloid Beta 1-40 BBB Shuttle Antibody, Clone 2H6

Figure 1 is a graph showing epitope mapping of antibody 2H6 by peptide competition. A(31_40 peptide immobilized on SA chip. Monoclonal antibodies 2289, 2286 or 2H6 (100 inM each), each pre-incubated with 16 gM for 1 hr

Various peptides (amino acids 1-16, 1-28, 1-38, 1-40, 1-42, 1-43, 17-40, 17-42, 22-35, 25-35 or 33-40 A( 3) Or no peptide flows to the chip.Measure the binding of antibody to immobilized A (31_40 peptide).

Figure 2 is a graph showing the binding of antibodies 2H6, 2286 and 2289 to different A (3 peptide C-terminal variants). GST-A (3 variants (M35A, V36A, G37A, G38A, V39A, or V40A), or GST-A (3 peptides 1-39, 1-41, 1-40, 1-42) were immobilized on ELISA plates.

Monoclonal antibodies 2286, 2H6 or 2289 (0.3 nM per mAb) were incubated with each immobilized peptide and their binding was detected by further incubation with biotinylated anti-mouse IgG.

Figure 3 is a graph showing that spatial learning deficits in APP transgenic mice were reversed after 16 weeks of antibody treatment with 2H6 and deglycosylated 2H6.

Mice were tested in a two-day radial arm water maze. The Y-axis represents the average number of mistakes made during the 2-day trial period. Block numbers 1-5 represent Day 1 testing; block numbers 6-10 represent Day 2 testing.

Figures 4A and 4B are graphs showing substantial Congo red-stained amyloid-beta peptide reduction in the hippocampus (Figure 4B) and frontal cortex (Figure 6A) after 16 weeks of treatment with the 2H6 anti-AMN antibody (designated AMN) , Deglycosylated 2H6 (D-2H6 for short) antibody. The Y-axis in Figures 4A and 4B represents the mean value of the percentage of areas positive for Congo red staining. The X-axis represents the type of antibody administered.

Figures 5A and 5B are shown using 2H6 anti-AMN (referred to as AMN), deglycosylated 2H6 (referred to as D-2H6) antibody. The Y-axis represents the mean of the percentage of areas positive for Congo red staining. The X-axis represents the type of antibody administered.

Figure 6 is a graph showing the number of Prussian blue positive profiles after 16 weeks of treatment with 2116, anti-AMN (designated AMN) and deglycosylated 2116 (designated D-2H6) antibodies. The Y-axis represents the normal section of each section. The X-axis represents the type of antibody administered.

Figure 7 is the serum A (3-peptide) level diagram after administration of APP Tg2576 mouse anti-AMN antibody (abbreviated AMN), antibody 2116 (abbreviated 2H6), deglycosylated 2H6 (abbreviated 2H6-D), and the levels in wild type (abbreviated as 2H6-D) After administration of anti-AMN antibody and antibody 2H6 in WT) mice.

Figure 8 shows immunostaining of CD45 in mouse hippocampus following intracranial administration of 2H6 antibody (A) or deglycosylated 2116 antibody (B).

Bottom panels show the ratio of the average area of injected versus non-injected sides to CD45-positive staining in the frontal cortex and hippocampus after intracranial administration of control antibody, 2H6 antibody, or deglycosylated 2116 antibody.

Figure 9 shows immunostaining of Fcy receptors in mouse hippocampus following intracranial administration of 2H6 antibody (A) or deglycosylated 2116 antibody (B).

The lower panel shows the ratio of the mean area occupied by Fcy receptor positive staining in the frontal cortex and hippocampus in the frontal cortex and hippocampus after intracranial administration of control antibody, 2116 antibody, or deglycosylated 2H6 antibody.

Figure 10 shows immunostaining of A(3 peptide) in mouse hippocampus following intracranial administration of 2H6 antibody (A) or deglycosylated 2H6 antibody (B).

Bottom panel shows the ratio of the mean area occupied by A(3 positive staining in the frontal cortex and hippocampus after intracranial administration of control antibody, 2H6 antibody, or deglycosylated 2H6 antibody on the injected side versus the non-injected side. "**" Indicates P<0.01 compared with the control antibody. "*" indicates P<0.05 compared with the control antibody.

Figure 11 shows Thioflavin-S in mouse hippocampus following intracranial administration of 2H6 antibody (A) or deglycosylated 2H6 antibody (B).

The lower panel shows the ratio of the mean area occupied by thioflavin-S positive staining in the frontal cortex and hippocampus in the frontal cortex and hippocampus after intracranial administration of control antibody, 2H6 antibody, or deglycosylated 2H6 antibody.

Figure 12 shows the epitope mapping of antibodies 2294 and 6G by ELISA.

Various A13 peptides were immobilized on ELISA plates. Antibodies were incubated with various immobilized peptides for 1 hour. Antibody 6G bound to immobilized A (measured with 3 peptides using goat anti-human kappa HRP-conjugated secondary antibody). Antibody 2294 bound to immobilized A (measured with goat anti-mouse antibody bound to heavy and light chains 3 peptides and are HRP-conjugated secondary antibodies. "NB" means no binding detected. Numbers in the column under "2294" and "6G" indicate absorbance at 450nm.