title: Novel Monoclonal Antibodies Recognizing Human Prostate-Specific Membrane Antigen (PSMA) as Research and Theranostic Tools.
abstract: 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.
authors.affiliation: Laboratory of Structural Biology, Institute of Biotechnology, Czech Academy of Sciences, Vestec, Czech Republic.
authors.forename: Zora
authors.initials: Z
authors.lastname: Nováková
authors.affiliation: The Russell H. Morgan Department of Radiology and Radiological Science, Johns Hopkins Medical Institutions, Baltimore, Maryland.
authors.forename: Catherine A
authors.initials: CA
authors.lastname: Foss
authors.affiliation: The Russell H. Morgan Department of Radiology and Radiological Science, Johns Hopkins Medical Institutions, Baltimore, Maryland.
authors.forename: Benjamin T
authors.initials: BT
authors.lastname: Copeland
authors.affiliation: Munich Center for Integrated Protein Science (CIPS-M) and Lehrstuhl für Biologische Chemie, Technische Universität München, Freising-Weihenstephan, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Morath
authors.affiliation: Laboratory of Structural Biology, Institute of Biotechnology, Czech Academy of Sciences, Vestec, Czech Republic.
authors.forename: Petra
authors.initials: P
authors.lastname: Baranová
authors.affiliation: Laboratory of Structural Biology, Institute of Biotechnology, Czech Academy of Sciences, Vestec, Czech Republic.
authors.forename: Barbora
authors.initials: B
authors.lastname: Havlínová
authors.affiliation: Munich Center for Integrated Protein Science (CIPS-M) and Lehrstuhl für Biologische Chemie, Technische Universität München, Freising-Weihenstephan, Germany.
authors.forename: Arne
authors.initials: A
authors.lastname: Skerra
authors.affiliation: The Russell H. Morgan Department of Radiology and Radiological Science, Johns Hopkins Medical Institutions, Baltimore, Maryland.
authors.forename: Martin G
authors.initials: MG
authors.lastname: Pomper
authors.affiliation: Laboratory of Structural Biology, Institute of Biotechnology, Czech Academy of Sciences, Vestec, Czech Republic.
authors.forename: Cyril
authors.identifier: http://orcid.org/0000-0003-2751-3060
authors.initials: C
authors.lastname: Barinka
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology; D058846:Antibodies, Monoclonal, Murine-Derived / Q000276:immunology* / Q000494:pharmacology; D000954:Antigens, Surface / Q000276:immunology; D043425:Glutamate Carboxypeptidase II / Q000037:antagonists & inhibitors / Q000276:immunology; D006801:Humans; D008297:Male; D051379:Mice; D011467:Prostate / Q000276:immunology / Q000473:pathology; D011471:Prostatic Neoplasms / Q000188:drug therapy / Q000276:immunology; D000068936:Theranostic Nanomedicine / Q000379:methods; D023041:Xenograft Model Antitumor Assays / Q000379:methods
chemical_list: D000911:Antibodies, Monoclonal; D058846:Antibodies, Monoclonal, Murine-Derived; D000954:Antigens, Surface; C478680:J591 monoclonal antibody; C473848:FOLH1 protein, human; D043425:Glutamate Carboxypeptidase II
keywords: NAALADase; glutamate carboxypeptidase II; in vivo imaging; monoclonal antibody; prostate cancer