Agroinfiltration of leaves for deconstructed viral vector-based transient gene expression: infiltrated leaf area affects recombinant hemagglutinin yield

Abstract Deconstructed viral vector systems for large-scale production of recombinant proteins in Nicotiana benthamiana plants require Agrobacterium tumefaciens-assisted delivery into mesophyll cells by vacuum infiltration of leaves. To clarify the importance of uniform infiltration over the leaf su...
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Gespeichert in:

Autor*in:	Matsuda, Ryo [verfasserIn] Kushibiki, Takanori [verfasserIn] Fujiuchi, Naomichi [verfasserIn] Fujiwara, Kazuhiro [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2018

Schlagwörter:	Hemagglutinin Influenza vaccine antigen Plant-made biopharmaceutical Vacuum infiltration

Übergeordnetes Werk:	Enthalten in: Horticulture, environment, and biotechnology - New York, NY : Springer, 2011, 59(2018), 4 vom: 05. Juni, Seite 547-555
Übergeordnetes Werk:	volume:59 ; year:2018 ; number:4 ; day:05 ; month:06 ; pages:547-555

Links:	Volltext

DOI / URN:	10.1007/s13580-018-0047-6

Katalog-ID:	SPR031863965

Internformat


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245	1	0	\|a Agroinfiltration of leaves for deconstructed viral vector-based transient gene expression: infiltrated leaf area affects recombinant hemagglutinin yield
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520			\|a Abstract Deconstructed viral vector systems for large-scale production of recombinant proteins in Nicotiana benthamiana plants require Agrobacterium tumefaciens-assisted delivery into mesophyll cells by vacuum infiltration of leaves. To clarify the importance of uniform infiltration over the leaf surface and to propose a possible method for uniform infiltration, we quantified the extent of leaf infiltration and evaluated the potential effect of uniform infiltration on recombinant protein yield. We also investigated the effects of plant characteristics (e.g., plant age, leaf dry mass per area) and leaf detachment treatment on the extent of infiltration. First, a simple method was developed to measure the extent of leaf infiltration using a red dye solution. The quantitative results showed that the extent of infiltration in young and old leaves was substantially lower than in mature leaves. However, recombinant hemagglutinin (HA), an influenza vaccine antigen, accumulated in the infiltrated area of young and old leaves, indicating that they can synthesize and accumulate HA at detectable levels. The extent of infiltration was affected by the plant age but not by leaf dry mass per area. Improving the extent of infiltration by supplemental syringe infiltration significantly increased total HA content in leaves. Thus, increasing the infiltrated leaf area represents a potential strategy for increasing the recombinant protein yield in deconstructed viral vector-based transient gene expression systems with A. tumefaciens. The extent of infiltration was also improved without the need for time-consuming syringe infiltration when detached leaves were subjected to vacuum infiltration, suggesting that this may be a potential method to increase the extent of infiltration.
650		4	\|a Hemagglutinin \|7 (dpeaa)DE-He213
650		4	\|a Influenza vaccine antigen \|7 (dpeaa)DE-He213
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650		4	\|a Vacuum infiltration \|7 (dpeaa)DE-He213
700	1		\|a Kushibiki, Takanori \|e verfasserin \|4 aut
700	1		\|a Fujiuchi, Naomichi \|e verfasserin \|4 aut
700	1		\|a Fujiwara, Kazuhiro \|e verfasserin \|4 aut
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912			\|a GBV_ILN_23
912			\|a GBV_ILN_24
912			\|a GBV_ILN_31
912			\|a GBV_ILN_32
912			\|a GBV_ILN_39
912			\|a GBV_ILN_40
912			\|a GBV_ILN_60
912			\|a GBV_ILN_62
912			\|a GBV_ILN_63
912			\|a GBV_ILN_65
912			\|a GBV_ILN_69
912			\|a GBV_ILN_70
912			\|a GBV_ILN_73
912			\|a GBV_ILN_74
912			\|a GBV_ILN_90
912			\|a GBV_ILN_95
912			\|a GBV_ILN_100
912			\|a GBV_ILN_105
912			\|a GBV_ILN_110
912			\|a GBV_ILN_120
912			\|a GBV_ILN_138
912			\|a GBV_ILN_150
912			\|a GBV_ILN_151
912			\|a GBV_ILN_161
912			\|a GBV_ILN_170
912			\|a GBV_ILN_171
912			\|a GBV_ILN_187
912			\|a GBV_ILN_213
912			\|a GBV_ILN_224
912			\|a GBV_ILN_230
912			\|a GBV_ILN_250
912			\|a GBV_ILN_281
912			\|a GBV_ILN_285
912			\|a GBV_ILN_293
912			\|a GBV_ILN_370
912			\|a GBV_ILN_602
912			\|a GBV_ILN_636
912			\|a GBV_ILN_702
912			\|a GBV_ILN_2001
912			\|a GBV_ILN_2003
912			\|a GBV_ILN_2004
912			\|a GBV_ILN_2005
912			\|a GBV_ILN_2006
912			\|a GBV_ILN_2007
912			\|a GBV_ILN_2008
912			\|a GBV_ILN_2009
912			\|a GBV_ILN_2010
912			\|a GBV_ILN_2011
912			\|a GBV_ILN_2014
912			\|a GBV_ILN_2015
912			\|a GBV_ILN_2020
912			\|a GBV_ILN_2021
912			\|a GBV_ILN_2025
912			\|a GBV_ILN_2026
912			\|a GBV_ILN_2027
912			\|a GBV_ILN_2031
912			\|a GBV_ILN_2034
912			\|a GBV_ILN_2037
912			\|a GBV_ILN_2038
912			\|a GBV_ILN_2039
912			\|a GBV_ILN_2044
912			\|a GBV_ILN_2048
912			\|a GBV_ILN_2049
912			\|a GBV_ILN_2050
912			\|a GBV_ILN_2055
912			\|a GBV_ILN_2057
912			\|a GBV_ILN_2059
912			\|a GBV_ILN_2061
912			\|a GBV_ILN_2064
912			\|a GBV_ILN_2065
912			\|a GBV_ILN_2068
912			\|a GBV_ILN_2070
912			\|a GBV_ILN_2086
912			\|a GBV_ILN_2088
912			\|a GBV_ILN_2093
912			\|a GBV_ILN_2106
912			\|a GBV_ILN_2107
912			\|a GBV_ILN_2108
912			\|a GBV_ILN_2110
912			\|a GBV_ILN_2111
912			\|a GBV_ILN_2112
912			\|a GBV_ILN_2113
912			\|a GBV_ILN_2116
912			\|a GBV_ILN_2118
912			\|a GBV_ILN_2119
912			\|a GBV_ILN_2122
912			\|a GBV_ILN_2129
912			\|a GBV_ILN_2143
912			\|a GBV_ILN_2144
912			\|a GBV_ILN_2147
912			\|a GBV_ILN_2148
912			\|a GBV_ILN_2152
912			\|a GBV_ILN_2153
912			\|a GBV_ILN_2188
912			\|a GBV_ILN_2232
912			\|a GBV_ILN_2336
912			\|a GBV_ILN_2446
912			\|a GBV_ILN_2470
912			\|a GBV_ILN_2472
912			\|a GBV_ILN_2507
912			\|a GBV_ILN_2522
912			\|a GBV_ILN_2548
912			\|a GBV_ILN_4035
912			\|a GBV_ILN_4037
912			\|a GBV_ILN_4046
912			\|a GBV_ILN_4112
912			\|a GBV_ILN_4125
912			\|a GBV_ILN_4242
912			\|a GBV_ILN_4246
912			\|a GBV_ILN_4249
912			\|a GBV_ILN_4251
912			\|a GBV_ILN_4305
912			\|a GBV_ILN_4306
912			\|a GBV_ILN_4307
912			\|a GBV_ILN_4313
912			\|a GBV_ILN_4322
912			\|a GBV_ILN_4323
912			\|a GBV_ILN_4324
912			\|a GBV_ILN_4325
912			\|a GBV_ILN_4326
912			\|a GBV_ILN_4333
912			\|a GBV_ILN_4334
912			\|a GBV_ILN_4335
912			\|a GBV_ILN_4336
912			\|a GBV_ILN_4338
912			\|a GBV_ILN_4393
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author_variant	r m rm t k tk n f nf k f kf
matchkey_str	article:22113460:2018----::gonitainfevsodcntutdiavcobsdrningnepesoiflrtdefra
hierarchy_sort_str	2018
publishDate	2018
allfields	10.1007/s13580-018-0047-6 doi (DE-627)SPR031863965 (SPR)s13580-018-0047-6-e DE-627 ger DE-627 rakwb eng 630 640 ASE Matsuda, Ryo verfasserin aut Agroinfiltration of leaves for deconstructed viral vector-based transient gene expression: infiltrated leaf area affects recombinant hemagglutinin yield 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Deconstructed viral vector systems for large-scale production of recombinant proteins in Nicotiana benthamiana plants require Agrobacterium tumefaciens-assisted delivery into mesophyll cells by vacuum infiltration of leaves. To clarify the importance of uniform infiltration over the leaf surface and to propose a possible method for uniform infiltration, we quantified the extent of leaf infiltration and evaluated the potential effect of uniform infiltration on recombinant protein yield. We also investigated the effects of plant characteristics (e.g., plant age, leaf dry mass per area) and leaf detachment treatment on the extent of infiltration. First, a simple method was developed to measure the extent of leaf infiltration using a red dye solution. The quantitative results showed that the extent of infiltration in young and old leaves was substantially lower than in mature leaves. However, recombinant hemagglutinin (HA), an influenza vaccine antigen, accumulated in the infiltrated area of young and old leaves, indicating that they can synthesize and accumulate HA at detectable levels. The extent of infiltration was affected by the plant age but not by leaf dry mass per area. Improving the extent of infiltration by supplemental syringe infiltration significantly increased total HA content in leaves. Thus, increasing the infiltrated leaf area represents a potential strategy for increasing the recombinant protein yield in deconstructed viral vector-based transient gene expression systems with A. tumefaciens. The extent of infiltration was also improved without the need for time-consuming syringe infiltration when detached leaves were subjected to vacuum infiltration, suggesting that this may be a potential method to increase the extent of infiltration. Hemagglutinin (dpeaa)DE-He213 Influenza vaccine antigen (dpeaa)DE-He213 Plant-made biopharmaceutical (dpeaa)DE-He213 Vacuum infiltration (dpeaa)DE-He213 Kushibiki, Takanori verfasserin aut Fujiuchi, Naomichi verfasserin aut Fujiwara, Kazuhiro verfasserin aut Enthalten in Horticulture, environment, and biotechnology New York, NY : Springer, 2011 59(2018), 4 vom: 05. Juni, Seite 547-555 (DE-627)680322655 (DE-600)2642577-4 2211-3460 nnns volume:59 year:2018 number:4 day:05 month:06 pages:547-555 https://dx.doi.org/10.1007/s13580-018-0047-6 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_105 GBV_ILN_110 GBV_ILN_120 GBV_ILN_138 GBV_ILN_150 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_187 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_250 GBV_ILN_281 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_636 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2031 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2039 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2070 GBV_ILN_2086 GBV_ILN_2088 GBV_ILN_2093 GBV_ILN_2106 GBV_ILN_2107 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2116 GBV_ILN_2118 GBV_ILN_2119 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2188 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2446 GBV_ILN_2470 GBV_ILN_2472 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_2548 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4393 GBV_ILN_4700 AR 59 2018 4 05 06 547-555
spelling	10.1007/s13580-018-0047-6 doi (DE-627)SPR031863965 (SPR)s13580-018-0047-6-e DE-627 ger DE-627 rakwb eng 630 640 ASE Matsuda, Ryo verfasserin aut Agroinfiltration of leaves for deconstructed viral vector-based transient gene expression: infiltrated leaf area affects recombinant hemagglutinin yield 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Deconstructed viral vector systems for large-scale production of recombinant proteins in Nicotiana benthamiana plants require Agrobacterium tumefaciens-assisted delivery into mesophyll cells by vacuum infiltration of leaves. To clarify the importance of uniform infiltration over the leaf surface and to propose a possible method for uniform infiltration, we quantified the extent of leaf infiltration and evaluated the potential effect of uniform infiltration on recombinant protein yield. We also investigated the effects of plant characteristics (e.g., plant age, leaf dry mass per area) and leaf detachment treatment on the extent of infiltration. First, a simple method was developed to measure the extent of leaf infiltration using a red dye solution. The quantitative results showed that the extent of infiltration in young and old leaves was substantially lower than in mature leaves. However, recombinant hemagglutinin (HA), an influenza vaccine antigen, accumulated in the infiltrated area of young and old leaves, indicating that they can synthesize and accumulate HA at detectable levels. The extent of infiltration was affected by the plant age but not by leaf dry mass per area. Improving the extent of infiltration by supplemental syringe infiltration significantly increased total HA content in leaves. Thus, increasing the infiltrated leaf area represents a potential strategy for increasing the recombinant protein yield in deconstructed viral vector-based transient gene expression systems with A. tumefaciens. The extent of infiltration was also improved without the need for time-consuming syringe infiltration when detached leaves were subjected to vacuum infiltration, suggesting that this may be a potential method to increase the extent of infiltration. Hemagglutinin (dpeaa)DE-He213 Influenza vaccine antigen (dpeaa)DE-He213 Plant-made biopharmaceutical (dpeaa)DE-He213 Vacuum infiltration (dpeaa)DE-He213 Kushibiki, Takanori verfasserin aut Fujiuchi, Naomichi verfasserin aut Fujiwara, Kazuhiro verfasserin aut Enthalten in Horticulture, environment, and biotechnology New York, NY : Springer, 2011 59(2018), 4 vom: 05. Juni, Seite 547-555 (DE-627)680322655 (DE-600)2642577-4 2211-3460 nnns volume:59 year:2018 number:4 day:05 month:06 pages:547-555 https://dx.doi.org/10.1007/s13580-018-0047-6 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_105 GBV_ILN_110 GBV_ILN_120 GBV_ILN_138 GBV_ILN_150 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_187 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_250 GBV_ILN_281 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_636 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2031 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2039 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2070 GBV_ILN_2086 GBV_ILN_2088 GBV_ILN_2093 GBV_ILN_2106 GBV_ILN_2107 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2116 GBV_ILN_2118 GBV_ILN_2119 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2188 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2446 GBV_ILN_2470 GBV_ILN_2472 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_2548 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4393 GBV_ILN_4700 AR 59 2018 4 05 06 547-555
allfields_unstemmed	10.1007/s13580-018-0047-6 doi (DE-627)SPR031863965 (SPR)s13580-018-0047-6-e DE-627 ger DE-627 rakwb eng 630 640 ASE Matsuda, Ryo verfasserin aut Agroinfiltration of leaves for deconstructed viral vector-based transient gene expression: infiltrated leaf area affects recombinant hemagglutinin yield 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Deconstructed viral vector systems for large-scale production of recombinant proteins in Nicotiana benthamiana plants require Agrobacterium tumefaciens-assisted delivery into mesophyll cells by vacuum infiltration of leaves. To clarify the importance of uniform infiltration over the leaf surface and to propose a possible method for uniform infiltration, we quantified the extent of leaf infiltration and evaluated the potential effect of uniform infiltration on recombinant protein yield. We also investigated the effects of plant characteristics (e.g., plant age, leaf dry mass per area) and leaf detachment treatment on the extent of infiltration. First, a simple method was developed to measure the extent of leaf infiltration using a red dye solution. The quantitative results showed that the extent of infiltration in young and old leaves was substantially lower than in mature leaves. However, recombinant hemagglutinin (HA), an influenza vaccine antigen, accumulated in the infiltrated area of young and old leaves, indicating that they can synthesize and accumulate HA at detectable levels. The extent of infiltration was affected by the plant age but not by leaf dry mass per area. Improving the extent of infiltration by supplemental syringe infiltration significantly increased total HA content in leaves. Thus, increasing the infiltrated leaf area represents a potential strategy for increasing the recombinant protein yield in deconstructed viral vector-based transient gene expression systems with A. tumefaciens. The extent of infiltration was also improved without the need for time-consuming syringe infiltration when detached leaves were subjected to vacuum infiltration, suggesting that this may be a potential method to increase the extent of infiltration. Hemagglutinin (dpeaa)DE-He213 Influenza vaccine antigen (dpeaa)DE-He213 Plant-made biopharmaceutical (dpeaa)DE-He213 Vacuum infiltration (dpeaa)DE-He213 Kushibiki, Takanori verfasserin aut Fujiuchi, Naomichi verfasserin aut Fujiwara, Kazuhiro verfasserin aut Enthalten in Horticulture, environment, and biotechnology New York, NY : Springer, 2011 59(2018), 4 vom: 05. Juni, Seite 547-555 (DE-627)680322655 (DE-600)2642577-4 2211-3460 nnns volume:59 year:2018 number:4 day:05 month:06 pages:547-555 https://dx.doi.org/10.1007/s13580-018-0047-6 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_105 GBV_ILN_110 GBV_ILN_120 GBV_ILN_138 GBV_ILN_150 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_187 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_250 GBV_ILN_281 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_636 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2031 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2039 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2070 GBV_ILN_2086 GBV_ILN_2088 GBV_ILN_2093 GBV_ILN_2106 GBV_ILN_2107 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2116 GBV_ILN_2118 GBV_ILN_2119 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2188 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2446 GBV_ILN_2470 GBV_ILN_2472 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_2548 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4393 GBV_ILN_4700 AR 59 2018 4 05 06 547-555
allfieldsGer	10.1007/s13580-018-0047-6 doi (DE-627)SPR031863965 (SPR)s13580-018-0047-6-e DE-627 ger DE-627 rakwb eng 630 640 ASE Matsuda, Ryo verfasserin aut Agroinfiltration of leaves for deconstructed viral vector-based transient gene expression: infiltrated leaf area affects recombinant hemagglutinin yield 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Deconstructed viral vector systems for large-scale production of recombinant proteins in Nicotiana benthamiana plants require Agrobacterium tumefaciens-assisted delivery into mesophyll cells by vacuum infiltration of leaves. To clarify the importance of uniform infiltration over the leaf surface and to propose a possible method for uniform infiltration, we quantified the extent of leaf infiltration and evaluated the potential effect of uniform infiltration on recombinant protein yield. We also investigated the effects of plant characteristics (e.g., plant age, leaf dry mass per area) and leaf detachment treatment on the extent of infiltration. First, a simple method was developed to measure the extent of leaf infiltration using a red dye solution. The quantitative results showed that the extent of infiltration in young and old leaves was substantially lower than in mature leaves. However, recombinant hemagglutinin (HA), an influenza vaccine antigen, accumulated in the infiltrated area of young and old leaves, indicating that they can synthesize and accumulate HA at detectable levels. The extent of infiltration was affected by the plant age but not by leaf dry mass per area. Improving the extent of infiltration by supplemental syringe infiltration significantly increased total HA content in leaves. Thus, increasing the infiltrated leaf area represents a potential strategy for increasing the recombinant protein yield in deconstructed viral vector-based transient gene expression systems with A. tumefaciens. The extent of infiltration was also improved without the need for time-consuming syringe infiltration when detached leaves were subjected to vacuum infiltration, suggesting that this may be a potential method to increase the extent of infiltration. Hemagglutinin (dpeaa)DE-He213 Influenza vaccine antigen (dpeaa)DE-He213 Plant-made biopharmaceutical (dpeaa)DE-He213 Vacuum infiltration (dpeaa)DE-He213 Kushibiki, Takanori verfasserin aut Fujiuchi, Naomichi verfasserin aut Fujiwara, Kazuhiro verfasserin aut Enthalten in Horticulture, environment, and biotechnology New York, NY : Springer, 2011 59(2018), 4 vom: 05. Juni, Seite 547-555 (DE-627)680322655 (DE-600)2642577-4 2211-3460 nnns volume:59 year:2018 number:4 day:05 month:06 pages:547-555 https://dx.doi.org/10.1007/s13580-018-0047-6 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_105 GBV_ILN_110 GBV_ILN_120 GBV_ILN_138 GBV_ILN_150 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_187 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_250 GBV_ILN_281 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_636 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2031 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2039 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2070 GBV_ILN_2086 GBV_ILN_2088 GBV_ILN_2093 GBV_ILN_2106 GBV_ILN_2107 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2116 GBV_ILN_2118 GBV_ILN_2119 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2188 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2446 GBV_ILN_2470 GBV_ILN_2472 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_2548 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4393 GBV_ILN_4700 AR 59 2018 4 05 06 547-555
allfieldsSound	10.1007/s13580-018-0047-6 doi (DE-627)SPR031863965 (SPR)s13580-018-0047-6-e DE-627 ger DE-627 rakwb eng 630 640 ASE Matsuda, Ryo verfasserin aut Agroinfiltration of leaves for deconstructed viral vector-based transient gene expression: infiltrated leaf area affects recombinant hemagglutinin yield 2018 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Abstract Deconstructed viral vector systems for large-scale production of recombinant proteins in Nicotiana benthamiana plants require Agrobacterium tumefaciens-assisted delivery into mesophyll cells by vacuum infiltration of leaves. To clarify the importance of uniform infiltration over the leaf surface and to propose a possible method for uniform infiltration, we quantified the extent of leaf infiltration and evaluated the potential effect of uniform infiltration on recombinant protein yield. We also investigated the effects of plant characteristics (e.g., plant age, leaf dry mass per area) and leaf detachment treatment on the extent of infiltration. First, a simple method was developed to measure the extent of leaf infiltration using a red dye solution. The quantitative results showed that the extent of infiltration in young and old leaves was substantially lower than in mature leaves. However, recombinant hemagglutinin (HA), an influenza vaccine antigen, accumulated in the infiltrated area of young and old leaves, indicating that they can synthesize and accumulate HA at detectable levels. The extent of infiltration was affected by the plant age but not by leaf dry mass per area. Improving the extent of infiltration by supplemental syringe infiltration significantly increased total HA content in leaves. Thus, increasing the infiltrated leaf area represents a potential strategy for increasing the recombinant protein yield in deconstructed viral vector-based transient gene expression systems with A. tumefaciens. The extent of infiltration was also improved without the need for time-consuming syringe infiltration when detached leaves were subjected to vacuum infiltration, suggesting that this may be a potential method to increase the extent of infiltration. Hemagglutinin (dpeaa)DE-He213 Influenza vaccine antigen (dpeaa)DE-He213 Plant-made biopharmaceutical (dpeaa)DE-He213 Vacuum infiltration (dpeaa)DE-He213 Kushibiki, Takanori verfasserin aut Fujiuchi, Naomichi verfasserin aut Fujiwara, Kazuhiro verfasserin aut Enthalten in Horticulture, environment, and biotechnology New York, NY : Springer, 2011 59(2018), 4 vom: 05. Juni, Seite 547-555 (DE-627)680322655 (DE-600)2642577-4 2211-3460 nnns volume:59 year:2018 number:4 day:05 month:06 pages:547-555 https://dx.doi.org/10.1007/s13580-018-0047-6 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_105 GBV_ILN_110 GBV_ILN_120 GBV_ILN_138 GBV_ILN_150 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_171 GBV_ILN_187 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_250 GBV_ILN_281 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_636 GBV_ILN_702 GBV_ILN_2001 GBV_ILN_2003 GBV_ILN_2004 GBV_ILN_2005 GBV_ILN_2006 GBV_ILN_2007 GBV_ILN_2008 GBV_ILN_2009 GBV_ILN_2010 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2015 GBV_ILN_2020 GBV_ILN_2021 GBV_ILN_2025 GBV_ILN_2026 GBV_ILN_2027 GBV_ILN_2031 GBV_ILN_2034 GBV_ILN_2037 GBV_ILN_2038 GBV_ILN_2039 GBV_ILN_2044 GBV_ILN_2048 GBV_ILN_2049 GBV_ILN_2050 GBV_ILN_2055 GBV_ILN_2057 GBV_ILN_2059 GBV_ILN_2061 GBV_ILN_2064 GBV_ILN_2065 GBV_ILN_2068 GBV_ILN_2070 GBV_ILN_2086 GBV_ILN_2088 GBV_ILN_2093 GBV_ILN_2106 GBV_ILN_2107 GBV_ILN_2108 GBV_ILN_2110 GBV_ILN_2111 GBV_ILN_2112 GBV_ILN_2113 GBV_ILN_2116 GBV_ILN_2118 GBV_ILN_2119 GBV_ILN_2122 GBV_ILN_2129 GBV_ILN_2143 GBV_ILN_2144 GBV_ILN_2147 GBV_ILN_2148 GBV_ILN_2152 GBV_ILN_2153 GBV_ILN_2188 GBV_ILN_2232 GBV_ILN_2336 GBV_ILN_2446 GBV_ILN_2470 GBV_ILN_2472 GBV_ILN_2507 GBV_ILN_2522 GBV_ILN_2548 GBV_ILN_4035 GBV_ILN_4037 GBV_ILN_4046 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4242 GBV_ILN_4246 GBV_ILN_4249 GBV_ILN_4251 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4326 GBV_ILN_4333 GBV_ILN_4334 GBV_ILN_4335 GBV_ILN_4336 GBV_ILN_4338 GBV_ILN_4393 GBV_ILN_4700 AR 59 2018 4 05 06 547-555
language	English
source	Enthalten in Horticulture, environment, and biotechnology 59(2018), 4 vom: 05. Juni, Seite 547-555 volume:59 year:2018 number:4 day:05 month:06 pages:547-555
sourceStr	Enthalten in Horticulture, environment, and biotechnology 59(2018), 4 vom: 05. Juni, Seite 547-555 volume:59 year:2018 number:4 day:05 month:06 pages:547-555
format_phy_str_mv	Article
institution	findex.gbv.de
topic_facet	Hemagglutinin Influenza vaccine antigen Plant-made biopharmaceutical Vacuum infiltration
dewey-raw	630
isfreeaccess_bool	false
container_title	Horticulture, environment, and biotechnology
authorswithroles_txt_mv	Matsuda, Ryo @@aut@@ Kushibiki, Takanori @@aut@@ Fujiuchi, Naomichi @@aut@@ Fujiwara, Kazuhiro @@aut@@
publishDateDaySort_date	2018-06-05T00:00:00Z
hierarchy_top_id	680322655
dewey-sort	3630
id	SPR031863965
language_de	englisch
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author	Matsuda, Ryo
spellingShingle	Matsuda, Ryo ddc 630 misc Hemagglutinin misc Influenza vaccine antigen misc Plant-made biopharmaceutical misc Vacuum infiltration Agroinfiltration of leaves for deconstructed viral vector-based transient gene expression: infiltrated leaf area affects recombinant hemagglutinin yield
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topic_title	630 640 ASE Agroinfiltration of leaves for deconstructed viral vector-based transient gene expression: infiltrated leaf area affects recombinant hemagglutinin yield Hemagglutinin (dpeaa)DE-He213 Influenza vaccine antigen (dpeaa)DE-He213 Plant-made biopharmaceutical (dpeaa)DE-He213 Vacuum infiltration (dpeaa)DE-He213
topic	ddc 630 misc Hemagglutinin misc Influenza vaccine antigen misc Plant-made biopharmaceutical misc Vacuum infiltration
topic_unstemmed	ddc 630 misc Hemagglutinin misc Influenza vaccine antigen misc Plant-made biopharmaceutical misc Vacuum infiltration
topic_browse	ddc 630 misc Hemagglutinin misc Influenza vaccine antigen misc Plant-made biopharmaceutical misc Vacuum infiltration
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title	Agroinfiltration of leaves for deconstructed viral vector-based transient gene expression: infiltrated leaf area affects recombinant hemagglutinin yield
ctrlnum	(DE-627)SPR031863965 (SPR)s13580-018-0047-6-e
title_full	Agroinfiltration of leaves for deconstructed viral vector-based transient gene expression: infiltrated leaf area affects recombinant hemagglutinin yield
author_sort	Matsuda, Ryo
journal	Horticulture, environment, and biotechnology
journalStr	Horticulture, environment, and biotechnology
lang_code	eng
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container_start_page	547
author_browse	Matsuda, Ryo Kushibiki, Takanori Fujiuchi, Naomichi Fujiwara, Kazuhiro
container_volume	59
class	630 640 ASE
format_se	Elektronische Aufsätze
author-letter	Matsuda, Ryo
doi_str_mv	10.1007/s13580-018-0047-6
dewey-full	630 640
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title_sort	agroinfiltration of leaves for deconstructed viral vector-based transient gene expression: infiltrated leaf area affects recombinant hemagglutinin yield
title_auth	Agroinfiltration of leaves for deconstructed viral vector-based transient gene expression: infiltrated leaf area affects recombinant hemagglutinin yield
abstract	Abstract Deconstructed viral vector systems for large-scale production of recombinant proteins in Nicotiana benthamiana plants require Agrobacterium tumefaciens-assisted delivery into mesophyll cells by vacuum infiltration of leaves. To clarify the importance of uniform infiltration over the leaf surface and to propose a possible method for uniform infiltration, we quantified the extent of leaf infiltration and evaluated the potential effect of uniform infiltration on recombinant protein yield. We also investigated the effects of plant characteristics (e.g., plant age, leaf dry mass per area) and leaf detachment treatment on the extent of infiltration. First, a simple method was developed to measure the extent of leaf infiltration using a red dye solution. The quantitative results showed that the extent of infiltration in young and old leaves was substantially lower than in mature leaves. However, recombinant hemagglutinin (HA), an influenza vaccine antigen, accumulated in the infiltrated area of young and old leaves, indicating that they can synthesize and accumulate HA at detectable levels. The extent of infiltration was affected by the plant age but not by leaf dry mass per area. Improving the extent of infiltration by supplemental syringe infiltration significantly increased total HA content in leaves. Thus, increasing the infiltrated leaf area represents a potential strategy for increasing the recombinant protein yield in deconstructed viral vector-based transient gene expression systems with A. tumefaciens. The extent of infiltration was also improved without the need for time-consuming syringe infiltration when detached leaves were subjected to vacuum infiltration, suggesting that this may be a potential method to increase the extent of infiltration.
abstractGer	Abstract Deconstructed viral vector systems for large-scale production of recombinant proteins in Nicotiana benthamiana plants require Agrobacterium tumefaciens-assisted delivery into mesophyll cells by vacuum infiltration of leaves. To clarify the importance of uniform infiltration over the leaf surface and to propose a possible method for uniform infiltration, we quantified the extent of leaf infiltration and evaluated the potential effect of uniform infiltration on recombinant protein yield. We also investigated the effects of plant characteristics (e.g., plant age, leaf dry mass per area) and leaf detachment treatment on the extent of infiltration. First, a simple method was developed to measure the extent of leaf infiltration using a red dye solution. The quantitative results showed that the extent of infiltration in young and old leaves was substantially lower than in mature leaves. However, recombinant hemagglutinin (HA), an influenza vaccine antigen, accumulated in the infiltrated area of young and old leaves, indicating that they can synthesize and accumulate HA at detectable levels. The extent of infiltration was affected by the plant age but not by leaf dry mass per area. Improving the extent of infiltration by supplemental syringe infiltration significantly increased total HA content in leaves. Thus, increasing the infiltrated leaf area represents a potential strategy for increasing the recombinant protein yield in deconstructed viral vector-based transient gene expression systems with A. tumefaciens. The extent of infiltration was also improved without the need for time-consuming syringe infiltration when detached leaves were subjected to vacuum infiltration, suggesting that this may be a potential method to increase the extent of infiltration.
abstract_unstemmed	Abstract Deconstructed viral vector systems for large-scale production of recombinant proteins in Nicotiana benthamiana plants require Agrobacterium tumefaciens-assisted delivery into mesophyll cells by vacuum infiltration of leaves. To clarify the importance of uniform infiltration over the leaf surface and to propose a possible method for uniform infiltration, we quantified the extent of leaf infiltration and evaluated the potential effect of uniform infiltration on recombinant protein yield. We also investigated the effects of plant characteristics (e.g., plant age, leaf dry mass per area) and leaf detachment treatment on the extent of infiltration. First, a simple method was developed to measure the extent of leaf infiltration using a red dye solution. The quantitative results showed that the extent of infiltration in young and old leaves was substantially lower than in mature leaves. However, recombinant hemagglutinin (HA), an influenza vaccine antigen, accumulated in the infiltrated area of young and old leaves, indicating that they can synthesize and accumulate HA at detectable levels. The extent of infiltration was affected by the plant age but not by leaf dry mass per area. Improving the extent of infiltration by supplemental syringe infiltration significantly increased total HA content in leaves. Thus, increasing the infiltrated leaf area represents a potential strategy for increasing the recombinant protein yield in deconstructed viral vector-based transient gene expression systems with A. tumefaciens. The extent of infiltration was also improved without the need for time-consuming syringe infiltration when detached leaves were subjected to vacuum infiltration, suggesting that this may be a potential method to increase the extent of infiltration.
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container_issue	4
title_short	Agroinfiltration of leaves for deconstructed viral vector-based transient gene expression: infiltrated leaf area affects recombinant hemagglutinin yield
url	https://dx.doi.org/10.1007/s13580-018-0047-6
remote_bool	true
author2	Kushibiki, Takanori Fujiuchi, Naomichi Fujiwara, Kazuhiro
author2Str	Kushibiki, Takanori Fujiuchi, Naomichi Fujiwara, Kazuhiro
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doi_str	10.1007/s13580-018-0047-6
up_date	2024-07-04T01:34:46.755Z
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fullrecord_marcxml	<?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01000caa a22002652 4500</leader><controlfield tag="001">SPR031863965</controlfield><controlfield tag="003">DE-627</controlfield><controlfield tag="005">20230519175353.0</controlfield><controlfield tag="007">cr uuu---uuuuu</controlfield><controlfield tag="008">201007s2018 xx \|\|\|\|\|o 00\| \|\|eng c</controlfield><datafield tag="024" ind1="7" ind2=" "><subfield code="a">10.1007/s13580-018-0047-6</subfield><subfield code="2">doi</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-627)SPR031863965</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(SPR)s13580-018-0047-6-e</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-627</subfield><subfield code="b">ger</subfield><subfield code="c">DE-627</subfield><subfield code="e">rakwb</subfield></datafield><datafield tag="041" ind1=" " ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="082" ind1="0" ind2="4"><subfield code="a">630</subfield><subfield code="a">640</subfield><subfield code="q">ASE</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Matsuda, Ryo</subfield><subfield code="e">verfasserin</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Agroinfiltration of leaves for deconstructed viral vector-based transient gene expression: infiltrated leaf area affects recombinant hemagglutinin yield</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="c">2018</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="a">Text</subfield><subfield code="b">txt</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="a">Computermedien</subfield><subfield code="b">c</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="a">Online-Ressource</subfield><subfield code="b">cr</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="520" ind1=" " ind2=" "><subfield code="a">Abstract Deconstructed viral vector systems for large-scale production of recombinant proteins in Nicotiana benthamiana plants require Agrobacterium tumefaciens-assisted delivery into mesophyll cells by vacuum infiltration of leaves. To clarify the importance of uniform infiltration over the leaf surface and to propose a possible method for uniform infiltration, we quantified the extent of leaf infiltration and evaluated the potential effect of uniform infiltration on recombinant protein yield. We also investigated the effects of plant characteristics (e.g., plant age, leaf dry mass per area) and leaf detachment treatment on the extent of infiltration. First, a simple method was developed to measure the extent of leaf infiltration using a red dye solution. The quantitative results showed that the extent of infiltration in young and old leaves was substantially lower than in mature leaves. However, recombinant hemagglutinin (HA), an influenza vaccine antigen, accumulated in the infiltrated area of young and old leaves, indicating that they can synthesize and accumulate HA at detectable levels. The extent of infiltration was affected by the plant age but not by leaf dry mass per area. Improving the extent of infiltration by supplemental syringe infiltration significantly increased total HA content in leaves. Thus, increasing the infiltrated leaf area represents a potential strategy for increasing the recombinant protein yield in deconstructed viral vector-based transient gene expression systems with A. tumefaciens. The extent of infiltration was also improved without the need for time-consuming syringe infiltration when detached leaves were subjected to vacuum infiltration, suggesting that this may be a potential method to increase the extent of infiltration.</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">Hemagglutinin</subfield><subfield code="7">(dpeaa)DE-He213</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">Influenza vaccine antigen</subfield><subfield code="7">(dpeaa)DE-He213</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">Plant-made biopharmaceutical</subfield><subfield code="7">(dpeaa)DE-He213</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">Vacuum infiltration</subfield><subfield code="7">(dpeaa)DE-He213</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Kushibiki, Takanori</subfield><subfield code="e">verfasserin</subfield><subfield code="4">aut</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Fujiuchi, Naomichi</subfield><subfield code="e">verfasserin</subfield><subfield code="4">aut</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Fujiwara, Kazuhiro</subfield><subfield code="e">verfasserin</subfield><subfield code="4">aut</subfield></datafield><datafield tag="773" ind1="0" ind2="8"><subfield code="i">Enthalten in</subfield><subfield code="t">Horticulture, environment, and biotechnology</subfield><subfield code="d">New York, NY : Springer, 2011</subfield><subfield code="g">59(2018), 4 vom: 05. 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Agroinfiltration of leaves for deconstructed viral vector-based transient gene expression: infiltrated leaf area affects recombinant hemagglutinin yield

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