Michael J Mullin1,2, Claire Wilkinson3,4, Ian Hiles5,4, Kathrine J Smith4. 1. Protein Technologies Group, GlaxoSmithKline Research & Development, Gunnels Wood Road, Stevenage, SG1 2NY, Hertfordshire, UK. Michael.J.Mullin@gsk.com. 2. Department of Protein & Cellular Sciences, GlaxoSmithKline Research & Development, Gunnels Wood Road, Stevenage, SG1 2NY, Hertfordshire, UK. Michael.J.Mullin@gsk.com. 3. Protein Technologies Group, GlaxoSmithKline Research & Development, Gunnels Wood Road, Stevenage, SG1 2NY, Hertfordshire, UK. 4. Department of Protein & Cellular Sciences, GlaxoSmithKline Research & Development, Gunnels Wood Road, Stevenage, SG1 2NY, Hertfordshire, UK. 5. Reagent Design Group, GlaxoSmithKline Research & Development, Gunnels Wood Road, Stevenage, SG1 2NY, Hertfordshire, UK.
Abstract
OBJECTIVE: To compare the effects of human Trypsin-1 signal peptide and pro-peptide on the expression and secretion efficiency of human Interleukin-25 from mammalian cells. RESULTS: The signal peptide and combined signal peptide-pro-peptide sequence of human Trypsin-1 improved the secretion of human IL-25 from 1.7 to 3.2 µg/ml and 1.7 to 8.2 µg/ml, respectively. Deletion analysis identified the minimal Trypsin-1 derived secretion domain that maintains improved human Interleukin-25 production and secretion. The presence of Trypsin-1 pro-peptide sequence does not affect the function of secreted human Interleukin-25. CONCLUSION: The Trypsin-1 signal peptide-pro-peptide sequence increased human IL-25 expression and secretion in mammalian cells by fivefold.
OBJECTIVE: To compare the effects of humanTrypsin-1 signal peptide and pro-peptide on the expression and secretion efficiency of humanInterleukin-25 from mammalian cells. RESULTS: The signal peptide and combined signal peptide-pro-peptide sequence of humanTrypsin-1 improved the secretion of humanIL-25 from 1.7 to 3.2 µg/ml and 1.7 to 8.2 µg/ml, respectively. Deletion analysis identified the minimal Trypsin-1 derived secretion domain that maintains improved humanInterleukin-25 production and secretion. The presence of Trypsin-1pro-peptide sequence does not affect the function of secreted humanInterleukin-25. CONCLUSION: The Trypsin-1 signal peptide-pro-peptide sequence increased humanIL-25 expression and secretion in mammalian cells by fivefold.
Entities:
Keywords:
Interleukin-25; Mammalian expression system; Pro-peptide; Signal peptide
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