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Expression in Trichoplusia ni (High Five) insect cells and partial purification of a chimeric version of active human factor VIII

Nersissian, Aram and Tseng, Kathryn L. and Remley, Luke T. and Madiba, Sally and Long, Amanda M. and Vacura, Brittany N. and Nangiana, Inderjit and Whitelegge, Julian P. (2016) Expression in Trichoplusia ni (High Five) insect cells and partial purification of a chimeric version of active human factor VIII. Haemophilia, 22 (S4). p. 20. ISSN 1351-8216. http://resolver.caltech.edu/CaltechAUTHORS:20160811-083054070

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Abstract

Introduction/Objectives: Purification of recombinant Factor VIII from mammalian cells poses remarkable challenge, because of its suboptimal yield and significant1y high cost. Bacalovirus derived expression in insect cells is an appropiate system to overcome these limitations. However, several reports have concluded that Sf9 cells are not suitable for expression of structurally correct factor VIII (fVIII). We have developed a cDNA construct encoding chimeric, full-length human fVIII in which dispensable B-domain has been replaced with significantly shorter B-domain derived from its ortholog in Fugu rubripe. This cDNA was successfully applied for expression and purificacion of active fVIII protein in Trichoplusia ni insect cells, known as High Five (Invitrogen).


Item Type:Article
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http://dx.doi.org/10.1111/hae.12980DOIArticle
http://onlinelibrary.wiley.com/doi/10.1111/hae.12980/abstractPublisherArticle
Additional Information:© 2016 The Authors Haemophilia © 2016 John Wiley & Sons Ltd. Version of Record online: 11 Jul 2016. Special Issue: WFH 2016 World Congress Abstracts, Orlando, Florida, USA, July 24–28, 2016.
Record Number:CaltechAUTHORS:20160811-083054070
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20160811-083054070
Official Citation:(2016), Abstracts. Haemophilia, 22: 3–138. doi: 10.1111/hae.12980
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:69564
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:11 Aug 2016 17:42
Last Modified:11 Aug 2016 17:46

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