Figure 7.

Tumor-targeted delivery of nivolumab by Her2-AAV. RENCA-Her2/neu bearing BALB/c mice received two injections of Her2-AAV^Nivo with 3.6 × 10¹¹ genome copies each, either intravenously (A: black lines; B: gray bars) or intratumorally (A: dashed lines; B: striped bars). Mice that received only PBS served as mock control. After 10 days organs were explanted and sera taken. Nivolumab present in tumor and liver organ lysates as well as in sera of the mice was quantified by sandwich ELISA using plates coated with recombinant PD-1 and an IgG-Fc-specific detection antibody. (A) Raw data obtained by ELISA. Values obtained from PBS-injected animals were subtracted from the ELISA signals of mice that had received AAV vectors. (B) Amounts of nivolumab present in tumor, liver, and serum. Values were normalized to the total protein yields for the organ lysates and calculated as ng/ml for the sera [tumor: n = 2 technical replicates per mouse, liver: n = 3 technical replicates per mouse; serum: n = 3 technical replicates per mouse; mean ± SD; ns, not significant by unpaired t-test (n = 3 mice per organ and injection route; numbers mean individual animal)].