| Budget Amount *help |
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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| Outline of Final Research Achievements |
In this study, I selected WT1 as my target. Next, I isolated TCR-like antibody clones by combining magnet beads screening, peptide pulsed T2A24 cell screening and plate screening strategy and isolated twelve type of antibodies. After testing recognition specificity and affinity, I chose #213 as candidate clone for chimeric antigen receptor (CAR) therapy.
Next, I investigated whether it is safe for cancer therapy. In the first, I checked critical amino acids for #213 CAR recognition by alanine substitution. Among nine amino acids of WT1p235-243, 2M, 3T, 4W, 5N were shown to be critical for #213 CAR recognition. Next, I further tested amino acid substitution by similar ones. 2W, 3N, 3S, 4F and 5H were shown to be recognized by #213 CAR. After blast search, I chose 42 off-target candidate peptides. In them, two peptides, that were expressed in normal tissues, were shown to be recognized by #213 CAR.
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