Yuan QA, Simmons HH, Robinson MK, Russeva M, Marasco WA, Adams GPDevelopment of engineered antibodies specific for the Mullerian inhibiting substance type II receptor: a promising candidate for targeted therapy of ovarian cancer. Mol Cancer Ther 5(8): 2096-2105

Department of Medical Oncology, Fox Chase Cancer Center, 333 Cottman Avenue, Philadelphia, PA 19111, USA.
Molecular Cancer Therapeutics (Impact Factor: 5.68). 09/2006; 5(8):2096-105. DOI: 10.1158/1535-7163.MCT-06-0115
Source: PubMed
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.

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    • "Amhr2 expression is detectable in 2-day-old mouse ovaries, a time when there are no follicles larger than primordial, but Amhr2 has not been localized to a particular cell type (Durlinger et al., 2002a). Given that AMH has been shown to inhibit in vitro invasion and migration of epithelial ovarian cancer cell lines that express its receptor AMHR2 (Chang et al., 2011) and that AMHR2 localizes to the surface epithelium in human ovaries (Yuan et al., 2006), it is interesting to speculate that exogenous AMH could have inhibited the migration of cells from the ovarian surface epithelium in the study by Nilsson et al. (2011). These results also imply that AMH activity/ availability should be tightly restricted during primordial follicle assembly to prevent disruption of the breakdown process. "
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    • "The target Her2 extracellular domain (ECD) was expressed from stably-transfected HEK-293 cells and purified using immobilized metal affinity chromatography (IMAC) as previously described [31]. The anti-Her2 scFv, H3, was isolated from a naïve human scFv phage display library using techniques essentially as previously described [32]. Her2 ECD obtained as described above was coated onto a Maxisorp-Immunotube (NUNC, Denmark) at a concentration of 20 μg/mL in coating buffer (Bup-H carbonate bicarbonate buffer; Pierce) at 4°C, overnight. "
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    • "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). "
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