Journal: European Journal of Immunology
Article Title: Selective Depletion of Autoreactive Plasma Cells as a Novel Strategy to Treat Acetylcholine Receptor Antibody‐Positive Myasthenia Gravis
doi: 10.1002/eji.70166
Figure Lengend Snippet: Integrity assessment of anti‐hCD38/⍺ECD‐ACE coupled by SA/biotin. (A) Schematic structure of anti‐hCD38/⍺ECD‐ACE coupled by SA/Biotin. Integrity analysis of anti‐hCD38/⍺ECD‐ACE confirmed by: (B ) ELISA via coating with anti‐αECD (D6) and anti‐βECD (B3/2) antibodies and detection with anti‐human IgG and (C) in vitro labeling of 2 × 10 5 hCD38‐D6 cells with 5 µg anti‐hCD38/⍺ECD‐ACE at a final concentration of 50 µg/mL and detection of both components of antibody (anti‐hCD38 and SA) and antigen (⍺ECD and biotin) by flow cytometry. (D) Experimental design to demonstrate the in vivo labeling of hCD38‐B3/2 (GFP + ) cells by intravenous injection of 200 µg anti‐hCD38/⍺ECD‐ACE. (E) Histogram graphs representing the in vivo labeling of cells and detection of antibody and antigen parts in both spleen and bone marrow of hCD38‐B3/2 (GFP + ) transferred into NSG‐Hc 1 mice ( n = 3 per group). anti‐hCD38/⍺ECD‐ACE, anti‐hCD38/⍺ECD‐antibody‐mediated cytotoxicity engager; hCD38‐B3/2, human CD38 transfected into mouse anti‐human AChR βECD secreting B3/2 (GFP + ) hybridoma cells.
Article Snippet: ELISA plates (Sarstedt) were coated with mouse‐anti‐hAChR αECD (5 μg/mL; clone D6, DRFZ) or mouse‐anti‐hAChR βECD (5 μg/mL; clone B3/2, DRFZ), blocked with PBS/3% BSA, and incubated with the sample for 2 h at 37°C.
Techniques: Enzyme-linked Immunosorbent Assay, In Vitro, Labeling, Concentration Assay, Flow Cytometry, In Vivo, Injection, Transfection