Journal: Neurotherapeutics

Article Title: A Novel Fusion Protein, AChR-Fc, Ameliorates Myasthenia Gravis by Neutralizing Antiacetylcholine Receptor Antibodies and Suppressing Acetylcholine Receptor-Reactive B Cells

doi: 10.1007/s13311-016-0476-9

Figure Lengend Snippet: Binding of the acetylcholine receptor (AChR)-Fc fusion protein to antiacetylcholine antibody receptor. (A) Surface plasmon resonance–kinetic analysis of AChR-Fc binding to mAb35. AChR-Fc was immobilized on a sensor chip and reacted with 6.25 to 200 nM of mAb35 at a temperature of 25 °C in 20 μl/ml solution. (B) Enzyme-linked immunosorbent analysis of the interaction between patient IgG and AChR-Fc. The IgG fraction purified from blood [patient(◆) or control(▲)] was immobilized at a concentration ranging from 0.24 μg/ml to 4.0 mg/ml, and subsequently reacted with AChR-Fc. RU = relative units

Article Snippet: Rat anti-AChR antibody mAb35 was prepared by using rat anti-AChR α1 subunit antibody-producing hybridoma cells (ATCC; TIB-175).

Techniques: Binding Assay, SPR Assay, Purification, Control, Concentration Assay

Journal: Neurotherapeutics

Article Title: A Novel Fusion Protein, AChR-Fc, Ameliorates Myasthenia Gravis by Neutralizing Antiacetylcholine Receptor Antibodies and Suppressing Acetylcholine Receptor-Reactive B Cells

doi: 10.1007/s13311-016-0476-9

Figure Lengend Snippet: Effectiveness of acetylcholine receptor (AChR)-Fc in a passive transfer myasthenia gravis (MG) model. Mean MG score of rats (6 in each group) administered an anti-AChR antibody (mAb35) and treated with AChR-Fc. Rats were treated 4 times with AChR-Fc (0.4, 2, 10 mg/kg). Mean and SEM are indicated. *p < 0.05 and **p < 0.01 compared with the control group

Article Snippet: Rat anti-AChR antibody mAb35 was prepared by using rat anti-AChR α1 subunit antibody-producing hybridoma cells (ATCC; TIB-175).

Techniques: Control

Journal: Frontiers in Immunology

Article Title: Discovery of functionally distinct anti-C7 monoclonal antibodies and stratification of anti-nicotinic AChR positive Myasthenia Gravis patients

doi: 10.3389/fimmu.2022.968206

Figure Lengend Snippet: (A) Schematic representation of Myasthenia Gravis disease biology driven by anti-AChR autoantibodies. (B–F) AF647 α-Bungarotoxin (in red) and nuclear stain (in blue) binding to cells transfected with AChRα1β1δϵ+Rapsyn (B) , AChRα1β1δϵ (C) , AChRα1β1 (D) , AChRδϵ (E) , Untransfected control cells (F, G) IgG, IgA, and IgM quantification of immunoglobulin-depleted vs. non-depleted NHS. (H) Haemolytic activity of immunoglobulin-depleted vs. non-depleted NHS. (I) Binding profile (IgG+IgA+IgM binding) of non-depleted NHS to transfected cells (blue: nuclear stain; red: AF647 anti-human IgG+IgA+IgM). (J) Binding profile (IgG+IgA+IgM binding) of immunoglobulin (Ig)-depleted NHS to transfected cells (blue: nuclear stain; red: AF647 anti-human IgG+IgA+IgM). (K) AChR decrease in AChR+Rapsyn-transfected cells incubated with tool anti-AChR antibody mAb35 in the presence of Ig-depleted NHS (n = 30) (One-way ANOVA (GraphPad Prism), Tukey’s multiple comparisons test). (L) Increased MAC deposition in AChR positive cells, incubated with tool anti-AChR antibody mAb35 in the presence of Ig-depleted NHS (n = 30) (One-way ANOVA (GraphPad Prism), Tukey’s multiple comparisons test). All NHS used and referred to in the remainder of the paper is Ig-depleted NHS. ****p <0.0001.

Article Snippet: A 10μg/ml mAb35 (Rat anti-AChR tool Ab, Genetex) solution was also prepared in culture medium.

Techniques: Staining, Binding Assay, Transfection, Control, Activity Assay, Incubation

Journal:

Article Title: Differential intra-endothelial delivery of polymer nanocarriers targeted to distinct PECAM-1 epitopes

doi: 10.1016/j.jconrel.2008.06.007

Figure Lengend Snippet: Monoclonal antibodies (mAb) recognizing distinct extracellular PECAM-1 epitopes

Article Snippet: Antibodies and reagents The mouse monoclonal antibodies to human PECAM-1 (mAbs) used in this study were mAb62 and mAb35 (kindly provided by Dr. M. Nakada, Centocor, Malvern, PA), mAbGi34 (AXXORA Platform, San Diego, CA), mAb4G6 and mAb37 [ 25 ].

Techniques: Inhibition

Journal:

Article Title: Differential intra-endothelial delivery of polymer nanocarriers targeted to distinct PECAM-1 epitopes

doi: 10.1016/j.jconrel.2008.06.007

Figure Lengend Snippet: Characterization of anti-PECAM/NCs

Article Snippet: Antibodies and reagents The mouse monoclonal antibodies to human PECAM-1 (mAbs) used in this study were mAb62 and mAb35 (kindly provided by Dr. M. Nakada, Centocor, Malvern, PA), mAbGi34 (AXXORA Platform, San Diego, CA), mAb4G6 and mAb37 [ 25 ].

Techniques:

Journal:

Article Title: Differential intra-endothelial delivery of polymer nanocarriers targeted to distinct PECAM-1 epitopes

doi: 10.1016/j.jconrel.2008.06.007

Figure Lengend Snippet: A: HUVECs were incubated at 4 °C in the presence of 0.13 µm or 5 µm diameter FITC-labeled carriers targeted by anti-PECAM mAb62 to permit carrier binding but not internalization. The cells were then washed and warmed to 37 °C either for 15 min, 1 h, or 3 h to allow internalization of pre-bound carriers. The cells were washed, fixed, and incubated with Texas red goat anti-mouse IgG which labels surface-bound carriers (yellow particles, arrowheads) vs internalized counterparts (green particles, arrows). Magnification bar= 10 µm. B: Quantification of the internalization kinetics of pre-bound mAb62 carriers of several sizes (0.13, 1 and 5 µm diameter) by HUVECs. C: Internalization of FITC-labeled carriers of various sizes (0.13, 1 and 5 µm diameter) targeted to PECAM-1 by mAb62, mAb35 or mAbGi34 after incubation with HUVEC for 1 h at 37 °C. Data in B and C are mean±SEM (n≥25 cells).

Article Snippet: Antibodies and reagents The mouse monoclonal antibodies to human PECAM-1 (mAbs) used in this study were mAb62 and mAb35 (kindly provided by Dr. M. Nakada, Centocor, Malvern, PA), mAbGi34 (AXXORA Platform, San Diego, CA), mAb4G6 and mAb37 [ 25 ].

Techniques: Incubation, Labeling, Binding Assay