306 Construction and characterization of a novel immunotoxin consisting of two ranpirnase (rpRNAse) molecules fused to an anti-CD74 humanized IgG4 antibody

Vanama, Sailaja; Chang, Chiung‐Wen; Sapra, Puja; Horak, Ivan D.; Hansen, Hinrich P.; Goldenberg, D.M.

doi:10.1016/s1359-6349(04)80314-8

Cited by 1 publication

(1 citation statement)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The selection of Rap(Q) over Rap as the fusion partner prevents the addition of carbohydrates to the fused L chain, which should render (Q)-hRS7 a more homogeneous product than 2L-Rap-hRS7 because only one band would be observed for the fused L chain on reducing SDS-PAGE. We also expect (Q)-hRS7 similar in vitro properties to 2L-Rap-hRS7 based on the results obtained for 2L-Rap-hLL1-γ4P and 2L-Rap(Q)-hLL1-γ4P (30).…”

Section: Introductionsupporting

confidence: 61%

Ranpirnase (Frog RNase) Targeted with a Humanized, Internalizing, Anti–Trop-2 Antibody Has Potent Cytotoxicity against Diverse Epithelial Cancer Cells

Chang

Gupta

Michel

et al. 2010

Molecular Cancer Therapeutics

View full text Add to dashboard Cite

Ranpirnase (Rap), an amphibian RNase, has been extensively studied both preclinically and clinically as an antitumor agent. Rap can be administered repeatedly to patients without any untoward immune response, with reversible renal toxicity reported to be dose limiting. To enhance its potency and targeted tumor therapy, we describe the generation of a novel IgG-based immunotoxin, designated 2L-Rap(Q)-hRS7, comprising Rap (Q), a mutant Rap with the putative N-glycosylation site removed, and hRS7, an internalizing, humanized antibody against Trop-2, a cell surface glycoprotein overexpressed in variety of epithelial cancers. The immunotoxin was generated recombinantly by fusing Rap(Q) to each of the two hRS7 light (L) chains at the NH 2 terminus, produced in stably transfected myeloma cells, purified by Protein A, and evaluated by a panel of in vitro studies. The results, including size-exclusion high-performance liquid chromatography, SDS-PAGE, flow cytometry, RNase activity, internalization, cell viability, and colony formation, showed its purity, molecular integrity, comparable affinity to hRS7 for binding to several Trop-2-expressing cell lines of different cancer types, and potency to inhibit growth of these cell lines at nanomolar concentrations. In addition, 2L-Rap (Q)-hRS7 suppressed tumor growth in a prophylactic model of nude mice bearing Calu-3 human non-small cell lung cancer xenografts, with an increase in the median survival time from 55 to 96 days (P < 0.01). These results warrant further development of 2L-Rap(Q)-hRS7 as a potential therapeutic for various Trop-2-expressing cancers, such as cervical, breast, colon, pancreatic, ovarian, and prostate cancers. Mol Cancer Ther; 9(8); 2276-86. ©2010 AACR.

show abstract

Section: Introductionsupporting

confidence: 61%