Patients with prostate-specific membrane antigen (PSMA)-positive
tumors can benefit from PSMA-targeted therapy; thus, we have constructed
a phage-displayed synthetic antibody library for the production of
novel PSMA antibodies with superior PSMA-targeting ability, favoring
clinical management. The binding affinities of anti-PSMA antibodies
were verified by an enzyme-linked immunosorbent assay (ELISA). Several
in vitro and in vivo experiments, including cellular uptake, internalization,
and cytotoxicity studies, micro single photon emission computed tomography
(microSPECT)/CT, and biodistribution studies, were performed to select
the most promising antibody among six different antibodies. The results
showed the target affinities of our antibodies in the ELISA assays
(7A, 8C, 8E, and 11A) were comparable to the existing antibodies (J591).
The half-maximal effective concentrations of 7A, 8C, 8E, 11A, and
J591 were 2.95, 6.64, 5.50, 2.08, and 4.79, respectively. The radiochemical
yield of
111
In-labeled antibodies ranged from 30% to 50%
with high radiochemical purity (>90%). In the cellular uptake studies,
the accumulated radioactivity of
111
In-J591,
111
In-7A, and
111
In-11A increased over time. The internalized
percentage of
111
In-11A was the highest (32.14% ±
2.06%) at 48 h after incubation, whereas that of
111
In-J591
peaked at 22.43% ± 4.38% at 24 h and dropped to 13.52% ±
3.03% at 48 h postincubation. Twenty-four hours after injection, radioactivity
accumulation appeared in the LNCaP xenografts of the mice injected
with
111
In-11A,
111
In-8E,
111
In-7A,
and
111
In-J591 but not in the xenografts of the
111
In-8C-injected group. Marked liver uptake was noticed in all groups
except the
111
In-11A-injected group. Moreover, the killing
effect of
177
Lu-11A was superior to that of
177
Lu-J591 at low concentrations. In conclusion, we successfully demonstrated
that 11A IgG owned the most optimal biological characteristics among
several new anti-PSMA antibodies and it can be an excellent PSMA-targeting
component for the clinical use.