Structures and antigenicities of truncated recombinant WT1 protein.

(A) Structures of the whole WT1 protein and of the His-tagged truncated recombinant WT1 protein HWT3 that was used as a WT1 antigen to detect WT1 antibodies. (B) SDS-PAGE analysis of truncated recombinant WT1 proteins. Lanes 1, 3, and 5: lysate of induced bacteria containing HWT2, HWT3, and HWT4, respectively. Lanes 2, 4, and 6: affinity-purified HWT2, HWT3, and HWT4, respectively. (C) Truncated recombinant WT1 proteins HWT3, HWT2, and HWT4 were reacted on a nitrocellulose membrane with anti-WT1 polyclonal antibody, WT180 (1), anti-WT1 monoclonal antibody against exon 5, 1B6 (2), or anti-His-tag antibodies (3), and dot spots were visualized as described in “Patients, materials, and methods.”