Production of recombinant endostatin from stably transformed Trichoplusia ni BTI Tn 5B1-4 cells |
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Authors: | Jong Min Lee Kyung Hwa Chang Jong Hwa Park Youn Hyung Lee In Sik Chung |
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Institution: | (1) Department of Genetic Engineering, Korea;(2) Plant Metabolism Research Center, Kyung Hee University, Suwon, 449-701, Korea |
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Abstract: | The recombinant plasmids harboring a heterologous gene coding mouse endostatin were transfected and expressed stably in Trichoplusia ni BTI Tn 5B1-4 (Tn 5B1-4) cells. Recombinant endostatin expressed in the stably transformed Tn 5B1-4 cells was secreted into the medium. Recombinant endostatin was also purified to homogeneity using a simple one-step Ni2+ affinity fractionation method. Purified recombinant endostatin inhibited endothelial cell proliferation in a dose-dependent manner. The concentration at half-maximum inhibition (ED50) for recombinant endostatin was approximately 0.35 g ml–1. In a T-flask, the stably transformed Tn 5B1-4 cells produced 14.3 mg recombinant endostatin l–1 at 6 days of cultivation. |
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Keywords: | in vitro activity mouse endostatin production Trichoplusia ni BTI Tn 5B1-4 cells |
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