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Bacterial Expression, Facile Purification and Properties of Recombinant Human Lymphotoxin (Tumor Necrosis Factor Beta)

Bio/Technology volume 7, pages 363–368 (1989)Cite this article

Abstract

We have expressed a synthetic recombinant human TNF–β(rhTNF–β) gene in Escherichia coli to give rhTNF–β in a soluble, highly active and intact form. The specific activity of rhTNF–β purified in a single, convenient step from bacterial lysates was 3–5 × 107 units/mg on mouse L–929 cells. The cytostatic (growth inhibitory) and cytotoxic (cell killing) activities of rhTNF–β were compared in a variety of untransformed and transformed cell lines. In the absence of the protein synthesis inhibitor cycloheximide, two cell lines were growth inhibited by rhTNF–β but none were lysed. In the presence of cycloheximide, five out of eleven cell lines became sensitive to lysis by rhTNF–β, indicating that some but not all cells require uninterrupted protein synthesis in order to protect themselves from TNF–β cytotoxicity. Cell lines varied greatly in their sensitivity to rhTNF–β and a different pattern of response was seen in the cytostatic and cytotoxic assays.

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  1. Heng-Fong Seow: Corresponding author

Authors and Affiliations

  1. Institute of Molecular and Cell Biology, National University of Singapore, Kent Ridge Crescent, Singapore, 0511, Republic of Singapore

    Heng-Fong Seow, Cynthia R. Goh, Lata Krishnan & Alan G. Porter

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Seow, HF., Goh, C., Krishnan, L. et al. Bacterial Expression, Facile Purification and Properties of Recombinant Human Lymphotoxin (Tumor Necrosis Factor Beta). Nat Biotechnol 7, 363–368 (1989). https://doi.org/10.1038/nbt0489-363

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