BACKGROUND: Prostate-specific membrane antigen (PSMA), also known as glutamate carboxypeptidase II (GCPII), is generally recognized as a diagnostic and therapeutic cancer antigen and a molecular address for targeted imaging and drug delivery studies. Due to its significance in cancer research, numerous monoclonal antibodies (mAbs) against GCPII have been described and marketed in the past decades. Unfortunately, some of these mAbs are poorly characterized, which might lead to their inappropriate use and misinterpretation of the acquired results. METHODS: We collected the 13 most frequently used mAbs against GCPII and quantitatively characterized their binding to GCPII by enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR). Using a peptide library, we mapped epitopes recognized by a given mAb. Finally, we assessed the applicability of these mAbs to routine experimental setups, including Western blotting, immunohistochemistry, and flow cytometry. RESULTS: ELISA and SPR analyses revealed that mAbs J591, J415, D2B, 107-1A4, GCP-05, and 2G7 bind preferentially to GCPII in native form, while mAbs YPSMA-1, YPSMA-2, GCP-02, GCP-04, and 3E6 bind solely to denatured GCPII. mAbs 24.4E6 and 7E11-C5.3 recognize both forms of GCPII. Additionally, we determined that GCP-02 and 3E6 cross-react with mouse GCPII, while GCP-04 recognizes GCPII and GCPIII proteins from both human and mouse. CONCLUSION: This comparative analysis provides the first detailed quantitative characterization of the most commonly used mAbs against GCPII and can serve as a guideline for the scientific community to use them in a proper and efficient way.

• Klíčová slova
• ELISA, NAALADase, Western blot, flow cytometry, folate hydrolase, glutamate carboxypeptidase II, immunohistochemistry, prostate-specific membrane antigen, surface plasmon resonance,
• MeSH
• adenokarcinom diagnóza   imunologie   metabolismus   MeSH
• ELISA MeSH
• imunohistochemie MeSH
• lidé MeSH
• mapování epitopu MeSH
• monoklonální protilátky analýza   imunologie   MeSH
• nádorové buněčné linie MeSH
• nádory prostaty diagnóza   imunologie   metabolismus   MeSH
• prostatický specifický antigen imunologie   metabolismus   MeSH
• průtoková cytometrie MeSH
• western blotting MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• monoklonální protilátky MeSH
• prostatický specifický antigen MeSH

Glutamate carboxypeptidase II (GCPII) and its splice variants, paralogs and human homologs represent a family of proteins with diverse tissue distribution, cellular localization and largely unknown function which have been explored only recently. While GCPII itself has been thoroughly studied from different perspectives, as clearly documented in this series of reviews, very little is known about other members of its family, even though they might be biologically relevant. Differential expression of individual GCPII splice variants is associated with tumor progression and prognosis of prostate cancer. The best studied GCPII homolog, GCPIII or NAALADase II, may be a valid pharmaceutical target for itself since it may compensate for a lack of normal GCPII enzymatic activity. Detailed molecular characterization of this family of proteins is thus very important not only with respect to the potential therapeutic use of GCPII inhibitors, but also for better understanding of the biological role of GCPII within as well as outside the nervous system.

• MeSH
• glutamátkarboxypeptidasa II analýza   antagonisté a inhibitory   genetika   metabolismus   MeSH
• inhibitory enzymů farmakologie   MeSH
• lidé MeSH
• molekulární sekvence - údaje MeSH
• protein - isoformy analýza   antagonisté a inhibitory   genetika   metabolismus   MeSH
• regulace genové exprese MeSH
• sekvence aminokyselin MeSH
• sekvenční seřazení MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• glutamátkarboxypeptidasa II MeSH
• inhibitory enzymů MeSH
• protein - isoformy MeSH

• MeSH
• atrézie střev chirurgie   MeSH
• jejunum abnormality   chirurgie   MeSH
• lidé MeSH
• mezenterium abnormality   MeSH
• novorozenec MeSH
• obstrukce duodena vrozené   chirurgie   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• novorozenec MeSH
• ženské pohlaví MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• kazuistiky MeSH