2016
DOI: 10.1002/biot.201500187
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Enhanced GAD65 production in plants using the MagnICON transient expression system: Optimization of upstream production and downstream processing

Abstract: Plants have emerged as competitive production platforms for pharmaceutical proteins that are required in large quantities. One example is the 65-kDa isoform of human glutamic acid decarboxylase (GAD65), a major autoimmune diabetes autoantigen that has been developed as a vaccine candidate for the primary prevention of diabetes. The expression of GAD65 in plants has been optimized but large-scale purification is hampered by its tendency to associate with membranes. We investigated the potential for large-scale … Show more

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Cited by 12 publications
(9 citation statements)
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“…More recently, an N-terminally truncated form of GAD65mut (Δ87GAD65mut) was expressed in N. benthamiana and tobacco ( N. tabacum ) to increase the solubility and stability of the protein. This form of the protein also accumulated at higher levels than its full-length counterpart when expressed in N. benthamiana using the magnICON ® system ( Merlin et al, 2016 ).…”
Section: Resultsmentioning
confidence: 99%
See 3 more Smart Citations
“…More recently, an N-terminally truncated form of GAD65mut (Δ87GAD65mut) was expressed in N. benthamiana and tobacco ( N. tabacum ) to increase the solubility and stability of the protein. This form of the protein also accumulated at higher levels than its full-length counterpart when expressed in N. benthamiana using the magnICON ® system ( Merlin et al, 2016 ).…”
Section: Resultsmentioning
confidence: 99%
“…The magnICON ® Tobacco mosaic virus 3′ module pICH31070, containing the GAD65mut and Δ87GAD65mut sequences, was prepared as previously described ( Avesani et al, 2014 ; Merlin et al, 2016 ). The final vectors pICH31070.Δ87GAD65mut, pICH31070.GAD65mut, and pICH7410.GFP (3′ modules), pICH20111 (5′ module) and pICH14011 (integrase module) were introduced into Agrobacterium tumefaciens strain GV3101 by electroporation.…”
Section: Methodsmentioning
confidence: 99%
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“…Merlin et al [59] compared the yields of optimized target proteins after downstream processing in three different plant expression systems and reported that the highest yields of an optimized variant protein were achieved in a transient expression system using the MagnICON vector. They hypothesized that a significant increase in the yield of the TOI could be achieved using transient expression.…”
Section: Transient Expression Systemsmentioning
confidence: 99%