Figure A3.
The binding potency analysis of anti-EphA2 Mabs on CL1-5 cell by flow cytometry. Mouse anti human EphA2 antibodies (3F7) was used to detect EphA2 on CL1-5 cells. The cells were harvested from the tissue-cultured flask with versene (EDTA-PBS) and washed with ice cold 1% FBS/PBS. The cells were incubated with antibody (0.0625 mg/3 × 105 cells) for 30 min on ice and washed with 1% FBS/PB. The cells were incubated with FITC-conjugated donkey anti-mouse IgG antibody (0.2 mg/3 × 105 cells) for another 30 min on ice and washed with 1% FBS/PBS. The fluorescence intensity was determined by BD FACSCalibur (BD Biosciences). Data were analyzed by software FlowJo 7.51. The result of the flow cytometry analysis shows antibody3F7 has the high affinity to bind with EphA2 on CL1-5 cells.