Figure 2. Bioactivity of CLDN18.2, hFc, Human (Cat. No.: Z03709) was validated by ELISA.
Immobilized anti-Claudin 18.2 antibody was at 1 μg/mL (100 μL/well). Dose response curve for CLDN18.2, hFc, Human with the EC50 of 17.60 ng/ml was determined by ELISA. CLDN18.2, hFc, Human was found to be an active membrane protein.
Figure 3. Cell-based binding assay with CHO-K1/CLDN 18.2 stable cell line (Cat. No.: M00916).
CHO-K1/CLDN18.2 cells were plated at 5x105 cells/well in 100 µL of PBS in 96-well plate, incubated with gradient concentration of IMAB-362 (anti-CLDN18.2 antibody) on ice for 1 hour. The supernatant was discarded and the cells were incubated with goat anti-human IgG antibody at 10 µg/mL on ice for 1 hour. The mean fluorescence intensity of APC (Mean APC-H) was analyzed in live cell gate by a flow cytometer .
Figure 4. Cell based ELISA with CHO-K1/CLDN 18.2 cell line (Cat. No.: M00916).
CHO-K1/CLDN18.2 cells were plated at 8x104 cells /well in 100 μl DMEM plus with 10% FBS in 96-well plate overnight. Then the cells were fixed with paraformaldehyde for 15 minutes. After washing, the cells were incubated with gradient concentration of IMAB-362 (anti-CLDN18.2 antibody) for 1 hour, and incubated with goat anti-human IgG (H&L) [HRP] antibody. After incubation with TMB substrate, OD450 was measured by a microplate reader.