Development of engineered antibodies specific for the Müllerian inhibiting substance type II receptor: a promising candidate for targeted therapy of ovarian cancer

Abstract

The Müllerian inhibiting substance type II receptor (MISIIR) is involved in Müllerian duct regression as part of the development of the male reproductive system. In adult females, MISIIR is present on ovarian surface epithelium and is frequently expressed on human epithelial ovarian cancer cells. Müllerian inhibiting substance has been found to be capable of inhibiting the growth of primary human ovarian cancer cells derived from ascites and ovarian cancer cell lines. This suggested to us that MISIIR could be an attractive target for antibody-based tumor targeting and growth inhibition strategies. Here, we describe the production of recombinant human MISIIR extracellular domain-human immunoglobulin Fc domain fusion proteins and their use as targets for the selection of MISIIR-specific human single-chain variable fragments (scFv) molecules from a human nonimmune scFv phage display library. The binding kinetics of the resulting anti-MISIIR scFv clones were characterized and two were employed as the basis for the construction of bivalent scFv:Fc antibody-based molecules. Both bound specifically to human ovarian carcinoma cells in flow cytometry assays and cross-reacted with mouse MISIIR. These results indicate that antibody-based constructs may provide a highly specific means of targeting MISIIR on human ovarian carcinoma cells for the purpose of diagnosing and treating this disease.

Anti-Müllerian hormone inhibits growth of AMH type II receptor-positive human ovarian granulosa cell tumor cells by activating apoptosis

“…The production of purified, fully cleaved AMH has been achieved only recently, 51 and with this AMH one would need only 1:1000 of the dose compared with partially cleaved AMH. Targeting AMHRII with activating antibodies 62 or small molecule agonists 23,59 presents another attractive option for supplementary treatment of AMHRII-positive advanced or recurrent GCTs. Nevertheless, the observed activation of apoptosis in GCTs may not necessarily involve AMHRII, given that the doses used in this study are more robust than implicated by the reported K d for AMH to AMHRII.…”

Targeting ErbB2 and ErbB3 with a bispecific single-chain Fv enhances targeting selectivity and induces a therapeutic effect in vitro

“…The “dead” D3 scFv was isolated from a human phage-display library (Yuan et al, 2006) and binds to an epitope formed by the junction between a biotinylated peptide and streptavidin. Binding, as measured by surface plasmon resonance (SPR) on a BIAcore1000 (BIAcore, Piscataway, NJ), requires the presence of both the peptide and SA; binding cannot be competed with either of the components alone.…”

“…Binding of the A5 and the ALM bs-scFv, to ErbB2 and ErbB3 extracellular domains (ECDs) was characterized by SPR using ErbB2 and ErbB3 ECDs (Horak et al, 2005) as target antigens and methods described previously (Yuan et al, 2006). ECDs were diluted to 10 μg/mL in 10mM sodium acetate pH 5.2 and approximately 200 RU of ECDs were immobilized onto CM5 sensor chips via NHS-ester chemistry.…”

Expression of Antibody Fragments with a ControlledN-Glycosylation Pattern and Induction of Endoplasmic Reticulum-Derived Vesicles in Seeds of Arabidopsis

“…Thus, homodimerization but not heterotetramerization is necessary for correct folding, which could allow for more efficient expression. In general, scFv-Fcs exhibit characteristics equivalent to their parent IgG (Shu et al, 1993;Powers et al, 2001;Cao et al, 2009) and have been tested for similar applications (Li et al, 2000;Yuan et al, 2006;Mori and Kim, 2008;De Lorenzo and D'Alessio, 2009;Olafsen et al, 2009;Riccio et al, 2009). …”