Gene targeting strategy for B-CAG-hCD20 EL4. The human full-length CD20 CDS region was inserted in the ROSA26 exon1 of Wild-Type EL4.
Protein expression analysis
The CD20 expression analysis in B-CAG-hCD20 EL4 cells by flow cytometry. Single cells were collected from wild-type EL4 and B-CAG-hCD20 EL4 cells, and analyzed by flow cytometry with species-specific anti-CD20 antibody. Human CD20 was exclusively detectable in B-CAG-hCD20 EL4 cells but not wild-type EL4 cells.
Tumor Growth Curve & Body Weight Changes
Subcutaneous xenograft tumor growth of B-CAG-hCD20 EL4 cells. B-CAG-hCD20 EL4 cells and wild-type EL4 cells (2×105) were subcutaneously implanted into the C57BL/6N mice (n=5).Tumor size and mice body weight were measured twice a week. (A) Tumor average volume ± SEM, (B) Mice body weight (Mean± SEM). Volume was expressed in mm3 using the formula: V=0.5a X b2, where a and b were the long and short diameters of the tumor, respectively. As shown in panel A, B-CAG-hCD20 EL4 were able to establish tumor in vivo and can be used for efficacy study.
Tumor expression detection sample information
Protein expression analysis in tumor cells
FMO Control 1: B-CAG-hCD20 EL4 tumor cells only stained hCD20 antibody;
FMO Control 2: B-CAG-hCD20 EL4 tumor cells only stained mCD20 antibody.
B-CAG-hCD20 EL4 cells were transplanted into C57BL/6N mice(n=5), and on 31 days post inoculation, certain volumes of tumors as was taken to detect the expression level of the cells. As Shown,human CD20 was highly expressed on the surface of tumor cells. B-CAG-hCD20 EL4 can be used for in vivo efficacy validation.