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dc.contributor.authorMarquez, Jorge
dc.contributor.authorDong, Jianping
dc.contributor.authorDong, Chun
dc.contributor.authorTian, Changsheng
dc.contributor.authorSerrero, Ginette
dc.date.accessioned2021-02-08T18:43:41Z
dc.date.available2021-02-08T18:43:41Z
dc.date.issued2021-01-27
dc.identifier.urihttp://hdl.handle.net/10713/14510
dc.description.abstractAntibody-drug conjugates (ADC) are effective antibody-based therapeutics for hematopoietic and lymphoid tumors. However, there is need to identify new targets for ADCs, particularly for solid tumors and cancers with unmet needs. From a hybridoma library developed against cancer cells, we selected the mouse monoclonal antibody 33B7, which was able to bind to, and internalize, cancer cell lines. This antibody was used for identification of the target by immunoprecipitation and mass spectrometric analysis, followed by target validation. After target validation, 33B7 binding and target positivity were tested by flow cytometry and western blot analysis in several cancer cell lines. The ability of 33B7 conjugated to saporin to inhibit in vitro proliferation of PTFRN positive cell lines was investigated, as well as the 33B7 ADC in vivo effect on tumor growth in athymic mice. All flow cytometry and in vitro internalization assays were analyzed for statistical significance using a Welsh’s T-test. Animal studies were analyzed using Two-Way Analysis of Variance (ANOVA) utilizing post-hoc Bonferroni analysis, and/or Mixed Effects analysis. The 33B7 cell surface target was identified as Prostaglandin F2 Receptor Negative Regulator (PTGFRN), a transmembrane protein in the Tetraspanin family. This target was confirmed by showing that PTGFRN-expressing cells bound and internalized 33B7, compared to PTGFRN negative cells. Cells able to bind 33B7 were PTGFRN-positive by Western blot analysis. In vitro treatment PTGFRN-positive cancer cell lines with the 33B7-saporin ADC inhibited their proliferation in a dose-dependent fashion. 33B7 conjugated to saporin was also able to block tumor growth in vivo in mouse xenografts when compared to a control ADC. These findings show that screening antibody libraries for internalizing antibodies in cancer cell lines is a good approach to identify new cancer targets for ADC development. These results suggest PTGFRN is a possible therapeutic target via antibody-based approach for certain cancers.en_US
dc.description.urihttps://doi.org/10.1371/journal.pone.0246197en_US
dc.language.isoenen_US
dc.publisherPublic Library of Scienceen_US
dc.relation.ispartofPLoS ONEen_US
dc.subjectantibody-drug conjugatesen_US
dc.subject.lcshCanceren_US
dc.subject.meshImmunoconjugatesen_US
dc.titleIdentification of Prostaglandin F2 Receptor Negative Regulator (PTGFRN) as an internalizable target in cancer cells for antibody-drug conjugate developmenten_US
dc.typeArticleen_US
dc.identifier.doi10.1371/journal.pone.0246197
dc.identifier.pmid33503070
dc.source.volume16
dc.source.issue1
dc.source.beginpagee0246197
dc.source.endpage
dc.source.countryUnited States


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