This work was supported by the COST action &#8216;Molecular pharming: Plants as a production platform for high-value proteins&#8217; FA0804. The Authors thank Dr Anatoli Giritch and Prof. Yuri Gleba for providing the MagnICON vectors for research purposes.

<ol>
	<li>Hampe, C. S., Hammerle, L. P., Falorni, A., Robertson, J., Lernmark, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Site-directed+mutagenesis+of+K396R+of+the+65+kDa+glutamic+acid+decarboxylase+active+site+obliterates+enzyme+activity+but+not+antibody+binding.">Site-directed mutagenesis of K396R of the 65 kDa glutamic acid decarboxylase active site obliterates enzyme activity but not antibody binding.</a> FEBS Lett. 488 (3), 185-189 (2001).</li><li>Avesani, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recombinant+human+GAD65+accumulates+to+high+levels+in+transgenic+tobacco+plants+when+expressed+as+an+enzymatically+inactive+mutant.">Recombinant human GAD65 accumulates to high levels in transgenic tobacco plants when expressed as an enzymatically inactive mutant.</a> Plant Biotechnol. J. 9 (8), 862-872 (2010).</li><li>Sambrook, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Molecular Cloning: A laboratory manual. Second Edition. , (1989).</li><li>Avesani, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+analysis+of+different+biofactories+for+the+production+of+a+major+diabetes+autoantigen.">Comparative analysis of different biofactories for the production of a major diabetes autoantigen.</a> Transgenic Res. 23, 281-291 (2014).</li><li>Marillonnet, S., Giritch, A., Gils, M., Kandzia, R., Klimyuk, V., Gleba, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+planta+engineering+of+viral+RNA+replicons:+efficient+assembly+by+recombination+of+DNA+modules+delivered+by+Agrobacterium.">In planta engineering of viral RNA replicons: efficient assembly by recombination of DNA modules delivered by Agrobacterium.</a> Proc. Natl. Acad. Sci. (USA). 101 (18), 6852-6857 (2004).</li><li>Gleba, Y., Klimyuk, V., Marillonnet, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Viral+vectors+for+the+expression+of+proteins+in+plants).">Viral vectors for the expression of proteins in plants).</a> Curr. Opin. Biotechnol. 18, 134-141 (2007).</li><li>Engler, C., Kandzia, R., Marillonnet, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+one+pot,+one+step,+precision+cloning+method+with+high+throughput+capability.">A one pot, one step, precision cloning method with high throughput capability.</a> PLoS One. 3 (11), (2008).</li><li>Xu, R., Li, Q. Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protocol:+streamline+cloning+of+genes+into+binary+vectors+in+Agrobacterium+via+the+Gateway+TOPO+vector+system.">Protocol: streamline cloning of genes into binary vectors in Agrobacterium via the Gateway TOPO vector system.</a> Plant Methods. 4 (4), 1-7 (2008).</li><li>Fairbanks, G., Steck, T. L., Wallach, D. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophoretic+analysis+of+the+major+polypeptides+of+the+human+erythrocyte+membrane.">Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane.</a> Biochemistry. 10 (13), 2606-2617 (1971).</li><li>Falorni, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radioimmunoassay+detects+the+frequent+occurrence+of+autoantibodies+to+the+Mr+65,000+isoform+of+glutamic+acid+decarboxylase+in+Japanese+insulin-dependent+diabetes.">Radioimmunoassay detects the frequent occurrence of autoantibodies to the Mr 65,000 isoform of glutamic acid decarboxylase in Japanese insulin-dependent diabetes.</a> Autoimmunity. 19, 113-125 (1994).</li><li>Hunt, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+gene+to+protein:+a+review+of+new+and+enabling+technologies+for+multi-parallel+protein+expression.">From gene to protein: a review of new and enabling technologies for multi-parallel protein expression.</a> Protein Expr. Purif. 40 (1), 1-22 (2005).</li><li>Arzola, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transient+co-expression+of+post-transcriptional+silencing+suppressor+for+increased+in+planta+expression+of+a+recombinant+anthrax+receptor+fusion+protein.">Transient co-expression of post-transcriptional silencing suppressor for increased in planta expression of a recombinant anthrax receptor fusion protein.</a> Int. J. Mol. Sci. 12 (8), 4975-4990 (2011).</li><li>Merlin, M., Gecchele, E., Capaldi, S., Pezzotti, M., Avesani, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+evaluation+of+recombinant+protein+production+in+different+biofactories:+the+green+perspective.">Comparative evaluation of recombinant protein production in different biofactories: the green perspective.</a> Biomed. Res. Int. 2014, 136419 (2014).</li><li>Avesani, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improved+in+planta+expression+of+the+human+islet+autoantigen+glutamic+acid+decarboxylase+(GAD65).">Improved in planta expression of the human islet autoantigen glutamic acid decarboxylase (GAD65).</a> Transgenic Res. 12 (2), 203-212 (2003).</li><li>Leuzinger, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+agroinfiltration+of+Plants+for+high-level+transient+expression+of+recombinant+proteins.">Efficient agroinfiltration of Plants for high-level transient expression of recombinant proteins.</a> J Vis Exp. (77), (2013).</li></ol>

The authors declare that there is no conflict of interests regarding the publication of this paper.

Bakteriyel hücreler, Bakulovirüs / böcek hücreleri ve bitkiler: Bu çalışmada üç farklı ortam, bir rekombinant insan proteininin sentezlenmesi için karşılaştırıldı. (- MagnICON ve pK7WG2 tabanlı - ve istikrarlı, yani geçici) bitki bazlı bir platform daha üç yaygın olarak kullanılan ifade teknolojilerini istismar tarafından araştırılmıştır. Bu deneyde, hGAD65mut için seçilen hedef protein önceden farklı sistemlerde 13 olarak ifade edildi ve üretim ve işlevselliği ...

Recombinant proteins are commercially mass-produced in heterologous expression systems with the aid of emerging biotechnology tools. Key factors that have to be considered when choosing the heterologous expression system include: protein quality, functionality, process speed, yield and cost.
In the recombinant protein field, the market for pharmaceuticals is expanding rapidly and, consequently, most biopharmaceuticals produced today are recombinant. Proteins can be expressed in cell cultures of bacteria, yeasts, molds, mammals, plants and insects, as well as in plant systems (either via stable- or transient-transformation) and transgenic animals; each expression system has its inherent advantages and limitations and for each target recombinant protein the optimal production system has to be carefully evaluated.
Plant-based platforms are arising as an important alternative to traditional fermenter-based systems for safe and cost-effective recombinant protein production. Although downstream processing costs are comparable to those of microbial and mammalian cells, the lower up-front investment required for commercial production in plants and the potential economy of scale, provided by cultivation over large areas, are key advantages.
We evaluated plants as bioreactors for the expression of the 65 kDa isoform of human glutamic acid decarboxylase (hGAD65), one of the major autoantigen in Type 1 autoimmune diabetes (T1D). hGAD65 is largely adopted as a marker, both for classifying and monitoring the progression of the disease and its role in T1D prevention is currently under investigation in clinical trials. If these trials are successful, the global demand for recombinant hGAD65 will increase dramatically.
Here, we focus on the expression of the enzymatically inactive counterpart of hGAD65, hGAD65mut, a mutant generated by substituting the lysine residue that binds the cofactor PLP (pyridoxal-5'-phosphate) with an arginine residue (K396R)1. 
hGAD65mut retains its immunogenicity and, in plant and insect cells, accumulates up to ten-fold higher than hGAD65, its wild-type counterpart. It was hypothesized that the enzymatic activity of hGAD65 interferes with plant cell metabolism to such an extent that it suppresses its own synthesis, whereas hGAD65mut, the enzymatically-inactive form, can be accumulated in plant cells to higher levels.
For the expression of hGAD65mut, the use of different technologies, widely used in plant biotechnology, was explored here and compared to traditional expression platforms (Escherichia coli and Baculovirus/insect cell-based).
In this work, the recombinant platforms developed for the expression of hGAD65mut comprising traditional and plant-based systems were reviewed and compared on the basis of process speed and yield, and of final product quality and functionality.

<table border="0" cellpadding="0" cellspacing="0" width="667">
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			<td dir="LTR" style="width:223px;">Do not freeze/thaw too many times</td>
		</tr>
		<tr height="40">
			<td dir="LTR" height="40" style="height:40px;width:191px;">SDS (Sodium dodecyl sulphate)</td>
			<td dir="LTR" style="width:111px;">Sigma</td>
			<td dir="LTR" style="width:143px;">L3771</td>
			<td dir="LTR" style="width:223px;">Flammable, toxic, corrosive &#8211; 10% stock</td>
		</tr>
		<tr height="20">
			<td dir="LTR" height="20" style="height:20px;width:191px;">Glycerol</td>
			<td dir="LTR" style="width:111px;">Sigma</td>
			<td dir="LTR" style="width:143px;">G5516</td>
			<td style="width:223px;"></td>
		</tr>
		<tr height="20">
			<td dir="LTR" height="20" style="height:20px;width:191px;">Brilliant Blue R-250</td>
			<td dir="LTR" style="width:111px;">Sigma</td>
			<td dir="LTR" style="width:143px;">B7920</td>
			<td style="width:223px;"></td>
		</tr>
		<tr height="20">
			<td dir="LTR" height="20" style="height:20px;width:191px;">Isopropanol</td>
			<td dir="LTR" style="width:111px;">Sigma</td>
			<td dir="LTR" style="width:143px;">24137</td>
			<td dir="LTR" style="width:223px;">Flammable</td>
		</tr>
		<tr height="20">
			<td dir="LTR" height="20" style="height:20px;width:191px;">Acetic acid</td>
			<td dir="LTR" style="width:111px;">Sigma</td>
			<td dir="LTR" style="width:143px;">27221</td>
			<td dir="LTR" style="width:223px;">Corrosive</td>
		</tr>
		<tr height="40">
			<td dir="LTR" height="40" style="height:40px;width:191px;">Anti-Glutamic acid decarboxylase 65/67</td>
			<td dir="LTR" style="width:111px;">Sigma</td>
			<td dir="LTR" style="width:143px;">G5163</td>
			<td dir="LTR" style="width:223px;">Do not freeze/thaw too many times</td>
		</tr>
		<tr height="60">
			<td dir="LTR" height="60" style="height:60px;width:191px;">Horseradish peroxidase (HRP)-conjugate anti-rabbit antibody</td>
			<td dir="LTR" style="width:111px;">Sigma</td>
			<td dir="LTR" style="width:143px;">A6154</td>
			<td dir="LTR" style="width:223px;">Do not freeze/thaw too many times</td>
		</tr>
		<tr height="40">
			<td dir="LTR" height="40" style="height:40px;width:191px;">Sf9 Cells</td>
			<td dir="LTR" style="width:111px;">Life Technologies</td>
			<td dir="LTR" style="width:143px;">11496</td>
			<td dir="LTR" style="width:223px;"></td>
		</tr>
		<tr height="40">
			<td dir="LTR" height="40" style="height:40px;width:191px;">BL21 Competent E.coli</td>
			<td dir="LTR" style="width:111px;">New England Biolabs</td>
			<td dir="LTR" style="width:143px;">C2530H</td>
			<td dir="LTR" style="width:223px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:191px;">Protein A Sepharose</td>
			<td style="width:111px;">Sigma</td>
			<td style="width:143px;">P2545</td>
			<td style="width:223px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:191px;">Cell culture plates&#160;</td>
			<td style="width:111px;">Sigma</td>
			<td style="width:143px;">CLS3516</td>
			<td style="width:223px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Radio Immuno Assay kit</td>
			<td>Techno Genetics</td>
			<td>12650805</td>
			<td style="width:223px;">Radioactive material&#160;</td>
		</tr>
	</tbody>
</table>

a comparative analysis of recombinant protein expression in different biofactories: bacteria, insect cells and plant systems

Bitki esaslı sistemler, yüksek kaliteli, biyolojik olarak aktif ürünler, esnek ve düşük maliyetli bir üretim için bunların iyi belgelenmiş potansiyel bir sonucu olarak, rekombinant proteinlerin üretimi için değerli bir platform olarak kabul edilir. Bu çalışmada, geçici ve stabil olarak bitki bazlı ekspresyon sistemleri, geleneksel fermentasyon cihazı dayalı hücre kültürlerinde (bakteriyel ve böcek) ve hedef insan yeniden birleştirici proteinin ifadesini karşılaştırıldı. Her platformda için, biz set-up, optimizasyon ve üretim sürecinin uzunluğu, nihai ürün kalitesi ve verimi açıklanan ve bizim seçtiğimiz hedef rekombinant protein için özel geçici üretim maliyetlerini, değerlendirildi. Genel olarak, sonuçlar bakteri bağlı erimez içerik korları dahilinde birikmesi, hedef proteinin üretimi için uygun olduğunu göstermektedir. Öte yandan, bitki bazlı sistemler, çok yönlü platformlar t&#39;ninşapka Bakulovirüs / böcek hücre sistemine daha düşük maliyetlerle seçilen proteinin üretimine izin verir. Özellikle, istikrarlı transgenik hatları nihai ürünün en yüksek verim ve geçici ifade bitkiler hızlı süreç geliştirme görüntülenen. Ancak, tüm rekombinant proteinleri bitki-temelli sistemlerde yararlanabilirler, ancak burada anlatıldığı gibi iyi üretim platformu, bir vaka-by-case yaklaşımı ile ampirik olarak tespit edilmelidir.

Farklı üretim sistemlerinde bir yeniden birleştirici hedef proteinin heterolog ekspresyonu için bir deney tasarımı burada tarif edilmektedir. ilk odak Her sistemde hedef proteinin ekspresyonu için uygun koşullar kurarak farklı düzlemlerin kurulum oldu. Hedef protein, hGAD65mut ekspresyonu, üçlü E. indüklenmiştir E. coli kültürleri. 37 ° C&#39;de ifade 3 saat sonra, bakteri hücreleri santrifüjleme ile toplandı ve sonikasyon ile parçalanır. Bir santrif?...

Watch this Scientific Journal Video about A Comparative Analysis of Recombinant Protein Expression in Different Biofactories: Bacteria, Insect Cells and Plant Systems at JoVE.com

A Comparative Analysis of Recombinant Protein Expression in Different Biofactories: Bacteria, Insect Cells and Plant Systems

In this study the expression of a target human recombinant protein in different production platforms was compared. We focused on traditional fermenter-based cultures and on plants, describing the set-up of each system and highlighting, on the basis of the reported results, the inherent limits and advantages for each platform.

Bakteriler, Böcek Hücreleri ve Bitki Sistemleri: Rekombinant Protein Farklı Biofactories İfade Karşılaştırmalı Analizi

Plant-based systems are considered a valuable platform for the production of recombinant proteins as a result of their well-documented potential for the ...

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23.2K Görüntüleme.  University of Verona, Verona, Italy. In this study the expression of a target human recombinant protein in different production platforms was compared. We focused on traditional fermenter-based cultures and on plants, describing the set-up of each system and highlighting, on the basis of the reported results, the inherent limits and advantages for each platform.