Detection of bystander effect in antibody-drug conjugates based on real-time label-free cell analysis technique
Objective To evaluate the bystander effect of antibody-drug conjugates with a real-time label-free cell analysis(RTCA).Methods RTCA was used to monitor the co-cultivation of Ag+(antigen-positive cells)and Ag-(antigen-negative cells)to observe the survival of Ag-cells.The experiment was repeated to validate the reliability and consistency of the results.Additionally,one-way ANOVA was used to compare the Ag-survival rates at a single time point among cultivation groups with different cell ratios.Results After adding antibody-drug conjugates drugs,the survival rate of Ag-decreased with the prolongation of co-culture time.In addition,at 96 h,compared to the Ag-cell survival rate with 0％Ag+cells,the Ag-cell survival rate of 10％Ag+cells was 89.07％(P=0.156),the Ag-cell survival rate with 25％Ag+cells was 68.93％(P=0.0026),and the Ag-cell survival rates with 50％Ag+,75％Ag+,and 90％Ag+cells were 35.28％,13.99％,and 12.02％,respectively(P＜0.0001).The bystander killing effect was significant within 25％～90％the proportion of Ag+cells.The repeatability test showed that the relative standard deviation of the bystander killing rates of different Ag+cells was less than 30％.Conclusion This method,which combines RTCA technology and co-cultivation,can more accurately simulate the bystander effect in biological processes,providing valuable reference for subsequent antibody-drug conjugate development.
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