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A Modified Human Tissue Plasminogen Activator with Extended Half–Life In Vivo

Bio/Technology volume 5, pages 953–958 (1987)Cite this article

An Erratum to this article was published on 01 June 1988

Abstract

We have produced a variant of human uterine tissue plasminogen activator (tPA) with increased in vivo fibrinolytic activity. The variant, termed mutant A, contains an asparagine to glutamine point mutation that prevents N–glycosylation at residue 451. Mutant A tPA was produced by in vitro mutagenesis of the tPA cDNA, followed by expression in mouse cells using a bovine papilloma virus (BPV) expression vector. After purification, the properties of mutant A and native tPA were compared. The two proteins have equivalent proteolytic activities and fibrin–binding properties, but mutant A tPA has a significantly longer systemic half–life in rabbits. In consequence, mutant A tPA has greater in vivo fibrinolytic activity than native tPA at an equivalent dose. These studies implicate glycosylation pattern as a significant determinant of systemic half–life of tPA and suggest that mutant A tPA will be an improved thrombolytic agent.

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Author notes

  1. Cha-Mer Wei

    Present address: Oncogen, Seattle, Washington, 98121

  2. David J. Livingston: To whom correspondence should be addressed.

Authors and Affiliations

  1. Integrated Genetics, Inc., Framingham, Massachusetts, 01701

    David Lau, Gregory Kuzma, Cha-Mer Wei, David J. Livingston & Nancy Hsiung

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Lau, D., Kuzma, G., Wei, CM. et al. A Modified Human Tissue Plasminogen Activator with Extended Half–Life In Vivo. Nat Biotechnol 5, 953–958 (1987). https://doi.org/10.1038/nbt0987-953

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