An affinity matured minibody for PET imaging of prostate stem cell antigen (PSCA)-expressing tumors.
OBJECTIVE
Prostate stem cell antigen (PSCA), a cell surface glycoprotein expressed in normal human prostate and bladder, is over-expressed in the majority of localized prostate cancer and most bone metastases. We have previously shown that the hu1G8 minibody, a humanized anti-PSCA antibody fragment (single-chain Fv-C(H)3 dimer, 80 kDa), can localize specifically and image PSCA-expressing xenografts at 21 h post-injection. However, the humanization and antibody fragment reformatting decreased its apparent affinity. Here, we sought to evaluate PET imaging contrast with affinity matured minibodies.


METHODS
Yeast scFv display, involving four rounds of selection, was used to generate the three affinity matured antibody fragments (A2, A11, and C5) that were reformatted into minibodies. These three affinity matured anti-PSCA minibodies were characterized in vitro, and following radiolabeling with (124)I were evaluated in vivo for microPET imaging of PSCA-expressing tumors.


RESULTS
The A2, A11, and C5 minibody variants all demonstrated improved affinity compared to the parental (P) minibody and were ranked as follows: A2 > A11 > C5 > P. The (124)I-labeled A11 minibody demonstrated higher immunoreactivity than the parental minibody and also achieved the best microPET imaging contrast in two xenograft models, LAPC-9 (prostate cancer) and Capan-1 (pancreatic cancer), when evaluated in vivo.


CONCLUSIONS
Of the affinity variant minibodies tested, the A11 minibody that ranked second in affinity was selected as the best immunoPET tracer to image PSCA-expressing xenografts. This candidate is currently under development for evaluation in a pilot clinical imaging study.
Lepin
Eric J
EJ
Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA 90095, USA. elepin@mednet.ucla.edu
Leyton
Jeffrey V
JV
Zhou
Yu
Y
Olafsen
Tove
T
Salazar
Felix B
FB
McCabe
Katelyn E
KE
Hahm
Scott
S
Marks
James D
JD
Reiter
Robert E
RE
Wu
Anna M
AM
D000595:Amino Acid Sequence; D000818:Animals; D000915:Antibody Affinity; D000951:Antigens, Neoplasm / Q000276:immunology* / Q000378:metabolism*; D045744:Cell Line, Tumor; D058851:GPI-Linked Proteins / Q000276:immunology / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007128:Immunoglobulin Fragments / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D009363:Neoplasm Proteins / Q000276:immunology* / Q000378:metabolism*; D049268:Positron-Emission Tomography / Q000379:methods*; D011471:Prostatic Neoplasms / Q000000981:diagnostic imaging* / Q000235:genetics* / Q000473:pathology; D011849:Radioactive Tracers
D000951:Antigens, Neoplasm; D058851:GPI-Linked Proteins; D007128:Immunoglobulin Fragments; D009363:Neoplasm Proteins; C110843:PSCA protein, human; D011849:Radioactive Tracers