BACKGROUND: Prostate-specific membrane antigen (PSMA) is a validated target for the imaging and therapy of prostate cancer. Here, we report the detailed characterization of four novel murine monoclonal antibodies (mAbs) recognizing human PSMA as well as PSMA orthologs from different species. METHODS: Performance of purified mAbs was assayed using a comprehensive panel of in vitro experimental setups including Western blotting, immunofluorescence, immunohistochemistry, ELISA, flow cytometry, and surface-plasmon resonance. Furthermore, a mouse xenograft model of prostate cancer was used to compare the suitability of the mAbs for in vivo applications. RESULTS: All mAbs demonstrate high specificity for PSMA as documented by the lack of cross-reactivity to unrelated human proteins. The 3F11 and 1A11 mAbs bind linear epitopes spanning residues 226-243 and 271-288 of human PSMA, respectively. 3F11 is also suitable for the detection of PSMA orthologs from mouse, pig, dog, and rat in experimental setups where the denatured form of PSMA is used. 5D3 and 5B1 mAbs recognize distinct surface-exposed conformational epitopes and are useful for targeting PSMA in its native conformation. Most importantly, using a mouse xenograft model of prostate cancer we show that both the intact 5D3 and its Fab fragment are suitable for in vivo imaging. CONCLUSIONS: With apparent affinities of 0.14 and 1.2 nM as determined by ELISA and flow cytometry, respectively, 5D3 has approximately 10-fold higher affinity for PSMA than the clinically validated mAb J591 and, therefore, is a prime candidate for the development of next-generation theranostics to target PSMA. Prostate 77:749-764, 2017. © 2017 Wiley Periodicals, Inc.

• Klíčová slova
• NAALADase, glutamate carboxypeptidase II, in vivo imaging, monoclonal antibody, prostate cancer,
• MeSH
• antigeny povrchové * imunologie   MeSH
• glutamátkarboxypeptidasa II * antagonisté a inhibitory   imunologie   MeSH
• lidé MeSH
• monoklonální protilátky imunologie   farmakologie   MeSH
• myší monoklonální protilátky imunologie   farmakologie   MeSH
• myši MeSH
• nádory prostaty * farmakoterapie   imunologie   MeSH
• prostata * imunologie   patologie   MeSH
• teranostická nanomedicína metody   MeSH
• xenogenní modely - testy protinádorové aktivity metody   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigeny povrchové * MeSH
• FOLH1 protein, human MeSH Prohlížeč
• glutamátkarboxypeptidasa II * MeSH
• J591 monoclonal antibody MeSH Prohlížeč
• monoklonální protilátky MeSH
• myší monoklonální protilátky MeSH

A highly purified 15 kDa glycoprotein isolated from ejaculated spermatozoa was used to raise antisera in female rabbits. An indirect immunofluorescence technique was used to detect the antigen in the seminal vesicle tissue and on the acrosomes of ejaculated, native and capacitated, boar spermatozoa. No immunoreactivity was detected on cells of the seminiferous tubules (spermatogonia, spermatocytes, and spermatids), on spermatozoa in the ductus epididymis and in cells of the epididymal and testicular tissues. These observations support the view that the 15 kDa protein is produced in the seminal vesicle secretory epithelium, and is attached to the sperm plasma membrane during the exposure of spermatozoa to seminal vesicle compounds. The observations that the antigen remained on the acrosome of ejaculated spermatozoa after capacitation and blocked sperm-oocyte binding in vitro suggest that the antigen plays a role in sperm-egg interactions. The strong immunoreactivity exhibited by cumulus cells after incubation of antisera with the porcine egg surrounded by cumulus cells shows the possible importance of the 15 kDa glycoprotein for contact of spermatozoa with cells of the cumulus oophorus surrounding the egg.

• MeSH
• aglutinace spermií MeSH
• ELISA MeSH
• epididymis imunologie   MeSH
• fluorescenční protilátková technika MeSH
• glykoproteiny imunologie   metabolismus   MeSH
• imunoelektroforéza MeSH
• interakce spermie a vajíčka imunologie   MeSH
• kapacitace spermií MeSH
• králíci MeSH
• ovariální folikul cytologie   metabolismus   MeSH
• prasata MeSH
• prostata imunologie   MeSH
• semenné váčky metabolismus   MeSH
• skot MeSH
• spermie imunologie   metabolismus   MeSH
• testis imunologie   MeSH
• zona pellucida metabolismus   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• mužské pohlaví MeSH
• skot MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• glykoproteiny MeSH

• MeSH
• Bacteria imunologie   MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• prostata imunologie   metabolismus   MeSH
• věkové faktory MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH