title: An albumin leader sequence coupled with a cleavage site modification enhances the yield of recombinant C-terminal Mullerian Inhibiting Substance.
abstract: Mullerian Inhibiting Substance (MIS) has been shown to inhibit ovarian cancer cells bothin-vitroandin-vivo. Furthermore, recent evidence suggests that MIS may effectively target a putative ovarian cancer progenitor cell population enriched by a panel of CD44+, CD24+, Ep-CAM+, and E-cadherin-cell surface markers. In order to accommodate clinical testing of MIS in ovarian cancer patients, the production of recombinant human MIS must be optimized to increase yield and purity. Here we show that, compared to wild type, the substitution of the MIS leader sequence to that of human serum albumin, combined with a modification of the endogenous cleavage site from RAQR/S to a furin/kex2 RARR/S consensus site results in high expression, increased C-terminus cleavage and a reduction in unwanted cryptic internal cleavage products when produced in CHO cells. Purified MIS containing these alterations retains its capacity to induce regression of the Mullerian duct in fetal rat embryonic urogenital ridge assays.
authors.affiliation: Pediatric Surgical Research Laboratories, Massachusetts General Hospital ; Harvard Medical School.
authors.forename: D
authors.initials: D
authors.lastname: Pépin
authors.affiliation: Pediatric Surgical Research Laboratories, Massachusetts General Hospital ; Harvard Medical School ; Massachusetts Eye-Ear Infirmary.
authors.forename: M
authors.initials: M
authors.lastname: Hoang
authors.affiliation: Pediatric Surgical Research Laboratories, Massachusetts General Hospital.
authors.forename: F
authors.initials: F
authors.lastname: Nicolaou
authors.affiliation: Pediatric Surgical Research Laboratories, Massachusetts General Hospital.
authors.forename: K
authors.initials: K
authors.lastname: Hendren
authors.affiliation: Pediatric Surgical Research Laboratories, Massachusetts General Hospital ; Harvard Medical School.
authors.forename: L A
authors.initials: LA
authors.lastname: Benedict
authors.affiliation: Harvard Medical School ; Massachusetts Eye-Ear Infirmary.
authors.forename: A
authors.initials: A
authors.lastname: Al-Moujahed
authors.affiliation: Pediatric Surgical Research Laboratories, Massachusetts General Hospital ; Harvard Medical School.
authors.forename: A
authors.initials: A
authors.lastname: Sosulski
authors.affiliation: Harvard Medical School ; Massachusetts Eye-Ear Infirmary.
authors.forename: A
authors.initials: A
authors.lastname: Marmalidou
authors.affiliation: Harvard Medical School ; Massachusetts Eye-Ear Infirmary.
authors.forename: D
authors.initials: D
authors.lastname: Vavvas
authors.affiliation: Pediatric Surgical Research Laboratories, Massachusetts General Hospital ; Harvard Medical School.
authors.forename: P K
authors.initials: PK
authors.lastname: Donahoe
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