Production and Characterization of Monoclonal Antibodies

The following primary mAbs were produced in our laboratories: M5A10 (IgG1, anti-PVR), L14 (IgG2a, anti-nectin-2), BAM195 (IgG1, anti-MICA), A6136 (IgM, anti-HLA class-I), NE97 (IgG2b, anti-B7-H3) (25 (link)), BAB281 (IgG1, anti-NKp46) (8 (link)). Anti-PD-L1.3.1 (IgG1, anti-PD-L1), and anti-PD-L2 (IgG1) were produced in D. Olive lab. D1.12 (IgG2a anti-HLA-II) mAb was produced by R. Accolla (University of Insubria, Varese, Italy). Anti-ULBP1-4 were purchased from R&D System (Minneapolis, US) and Santa Cruz Biotecnology (Dallas, US); anti-GD₂ (IgG2a) was purchased from BD Bioscience PharMingen, San Diego, CA); anti-CXCR4 (IgG2b) was purchased from &D Systems (Minneapolis, MN).
The following fluorochrome-conjugated anti-KIR in combination with anti-CD3-PECF594 (IgG1; BD Bioscience) and anti-CD56-PC7 (IgG1; Beckman Coulter, Marseille, France) mAbs were used: GL-183 (IgG1, anti-KIR2DL2/L3/S2; Beckman Coulter), EB6B (IgG1, anti-KIR2DL1/S1 and KIR2DL3*005; Beckman Coulter), Z27 (IgG1, anti-KIR3DL1/S1; Beckman Coulter), DX9 (IgG1, anti KIR3DL1; Miltenyi Biotec, Bergisch Gladbach, Germany), FES172 (IgG2a, KIR2DS4; Beckman Coulter), 143211 (IgG1, anti-KIR2DL1 and KIR2DS5; R&D). ECM41 mAb (IgM, anti-KIR2DL3, with the exception of *005; Our laboratory) followed by FITC-conjugated goat anti-mouse IgM (Southern Biotech, Birmingham, AL) was also used.