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  • Antibodies as Diagnostic Tools
    Nicotiana benthamiana leaves and Pichia pastoris De Meyer et al 2015 By making bivalent VHH Fc antibodies the VHH binding specificities are retained and sometimes augmented in terms of sensitivity by Fc based avidity effects and signal amplification with secondary anti Fc antibodies Moreover the Fc chain is a robust protein tag for purification and immunodetection and provides effector functions Currently together with the Stoger lab Vienna these VHH Fcs are being evaluated as immunomodulators in Arabidopsis thaliana seeds where they potentially interfere with antigen folding localization and or functioning in vivo References De Meyer T Eeckhout D De Rycke R De Buck S Muyldermans S and Depicker A 2014 Generation of VHH antibodies against the Arabidopsis thaliana seed storage proteins Plant Mol Biol 84 83 93 De Meyer T Laukens B Nolf J Van Lerberge E De Rycke R Debeuckelaer A De Buck S Callewaert N and Depicker A 2015 Comparison of VHH Fc antibody production in Arabidopsis thaliana Nicotiana benthamiana and Pichia pastoris Plant Biotechnol J in press Prev Next Projects Ann Depicker Group Members Lab Members and Alumni Publications Ann Depicker Research Groups Alain Goossens Lab Ann Depicker Lab Bruno Cammue Lab Daniel Van Damme Lab Dirk

    Original URL path: http://www.psb.vib-ugent.be/plant-made-antibodies/220-antibodies-as-diagnostic-tools (2016-04-26)
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  • Optimizing Seed-Based Production
    transformation with the constituting components For instance we have observed extreme difference in levels of assembled sSIgA in plants cotransformants with 3 T DNAs each encoding one of the components compared with one T DNA containing the the three constituting elements in tandem Further other factors like the influence of gene copy number of each component and the protein expression on the efficiency of assembly of sSIgA will be determined Aiming toward eventual valorization we are investigating the production of antibodies in the seeds of protein rich seed crop plants like soybean and pea In collaboration with the Lab of Inge Broer University of Rostock pea transformants were obtained and in the service facility of Kan Wang Iowa State University soya transformants were generated with 3 different antibody constructs Comparing different expression cassettes the possibilities and opportunities of each platform are being evaluated Further the seed produced antibodies in these different platforms are being characterized for their functionality proteolysis and glycosylation profile and the stability of the different formats of plant made antibodies are being studied in various conditions and environments It was already demonstrated that the recombinant antibodies stored in intact seeds are stable over several years Now a biochemical heat resistance profile on functionality will be made for purified antibodies and for antibodies in the seed matrix In parallel together with the Callewaert lab Unit of Medical Biotechnology VIB Ghent University we are developing novel strategies for glyco engineering in plants as the plant specific N glycans may represent immunogenic epitopes for humans and animals when used as injectables This is especially important when the seeds would be used to stockpile therapeutic recombinant antibodies that could be purified in emergencies like disease outbreaks cfr the ZMapp antibodies protecting against ebola Prev Next Projects Ann Depicker Group Members Lab

    Original URL path: http://www.psb.vib-ugent.be/plant-made-antibodies/222-optimizing-seed-based-production (2016-04-26)
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  • Protein Purification
    proteins of interest are isolated in a suitable buffer and with a desired purity and concentration All purifications are performed at 4 degrees C to preserve protein integrity Typical modes of protein purification include ion exchange hydrophobic interaction reversed phase size exclusion and affinity chromatography Often several techniques are performed consecutively to obtain the final product For example an initial crude enrichment step by affinity chromatography can be followed by ion exchange chromatography for further protein enrichment or by a size exclusion polishing step The process is highly dependant on the specific protein characteristics and hence requires customization Protein purification equipment ÄKTAexplorer and ÄKTApurifier system GE Healthcare Recommended Reading De Buck S Nolf J De Meyer T Virdi V De Wilde K Van Lerberge E Van Droogenbroeck B and Depicker A 2013 Fusion of an Fc chain to a VHH boosts the accumulation levels in Arabidopsis seeds Plant Biotechnol J 11 1006 1016 Virdi V Coddens A De Buck S Millet S Goddeeris B M Cox E De Greve H and Depicker A 2013 Orally fed seeds producing designer IgAs protect weaned piglets against enterotoxigenic Escherichia coli infection Proc Natl Acad Sci USA 110 11809 11814 Heijde M Binkert M Yin R H Ares Orpel F Rizzini L Van De Slijke E Persiau G Nolf J Gevaert K De Jaeger G and Ulm R 2013 Constitutively active UVR8 photoreceptor variant in Arabidopsis Proc Natl Acad Sci USA 110 20326 20331 Prev Projects Ann Depicker Group Members Lab Members and Alumni Publications Ann Depicker Research Groups Alain Goossens Lab Ann Depicker Lab Bruno Cammue Lab Daniel Van Damme Lab Dirk Inzé Lab Frank Van Breusegem Lab Geert De Jaeger Lab Ive De Smet Lab Jenny Russinova Lab Lieven De Veylder Lab Mieke Van Lijsebettens Lab Moritz Nowack Lab Sofie Goormachtig Lab

    Original URL path: http://www.psb.vib-ugent.be/plant-made-antibodies/219-protein-purification (2016-04-26)
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  • Bruno Cammue
    KU Leuven Centre of Microbial and Plant Genetics Kasteelpark Arenberg 20 box 2460 3001 Leuven Projects Bruno Cammue Publications Bruno Cammue Group Members Research Groups Alain Goossens Lab Ann Depicker Lab Bruno Cammue Lab Daniel Van Damme Lab Dirk Inzé Lab Frank Van Breusegem Lab Geert De Jaeger Lab Ive De Smet Lab Jenny Russinova Lab Lieven De Veylder Lab Mieke Van Lijsebettens Lab Moritz Nowack Lab Sofie Goormachtig Lab

    Original URL path: http://www.psb.vib-ugent.be/bruno-cammue (2016-04-26)
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  • Publications Bruno Cammue
    Members Research Groups Alain Goossens Lab Ann Depicker Lab Bruno Cammue Lab Daniel Van Damme Lab Dirk Inzé Lab Frank Van Breusegem Lab Geert De Jaeger Lab Ive De Smet Lab Jenny Russinova Lab Lieven De Veylder Lab Mieke Van Lijsebettens Lab Moritz Nowack Lab Sofie Goormachtig Lab Steven Maere Lab Tom Beeckman Lab Yves Van de Peer Lab Wout Boerjan Lab About PSB PSB Missions The Department Infrastructure Core

    Original URL path: http://www.psb.vib-ugent.be/publications-brunno-cammue (2016-04-26)
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  • Group Members
    Members Research Groups Alain Goossens Lab Ann Depicker Lab Bruno Cammue Lab Daniel Van Damme Lab Dirk Inzé Lab Frank Van Breusegem Lab Geert De Jaeger Lab Ive De Smet Lab Jenny Russinova Lab Lieven De Veylder Lab Mieke Van Lijsebettens Lab Moritz Nowack Lab Sofie Goormachtig Lab Steven Maere Lab Tom Beeckman Lab Yves Van de Peer Lab Wout Boerjan Lab About PSB PSB Missions The Department Infrastructure Core

    Original URL path: http://www.psb.vib-ugent.be/group-members-brcam (2016-04-26)
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  • Introduction
    Fungus Yeast Unit respectively Only the first unit is associated to the VIB Department of PSB and as such further described below The Fungus Yeast Unit studies the mode of action of bioactive peptides including PDFs and small molecules and uses yeast platforms as model systems for higher eukaryotes In our Plant Unit the discovery of different types of PDFs in Arabidopsis thaliana has significantly stimulated our research on their expression and function For example the discovery of the plant defensin AtPDF1 2a and the corresponding signaling pathways involved in its induction broadened the general insight in induced defense mechanisms in plants The corresponding gene is now world wide used as a marker for ethylene jasmonate controlled responses in plants To support this research different molecular tools and approaches have also been developed optimizing plant transformation and molecular breeding Current research is still focussing on 320 PDF like peptides in Arabidopsis using genome wide approaches such as the RNAseq technology primarily aiming at discovering their in planta role This research is recently broadened to other types of stress induced peptides SIPs Our expertise on resistance mechanisms mainly in A thaliana and focusing on resistance to necroptrophic fungi allowed us to successfully use this model plant as a source of resistance traits against agronomically important crops diseases e g Botrytis cinerea in tomato Fusarium oxysporum in banana Cercospora beticola in sugarbeet Cochliobolus heterostrophus in corn Based on the same expertise some years ago we also initiated research on defense mechanisms induced by biocontrol organisms the so called induced systemic response ISR This research is based on transcriptome studies both in a model plant A thaliana and in crops e g tomato lettuce involves both biotic and abiotic biocontrol agents and is mainly directed to different necroptrophic pathogens Bruno Cammue is a

    Original URL path: http://www.psb.vib-ugent.be/plant-fungi-interactions/225-introduction (2016-04-26)
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  • Daniel Van Damme
    B 9052 Gent BELGIUM Fax 32 0 9 33 13 809 Prev Next Projects Daniel Van Damme Group Members Lab Members and Alumni Group in Action Web Links Publications Daniel Van Damme Research Groups Alain Goossens Lab Ann Depicker Lab Bruno Cammue Lab Daniel Van Damme Lab Dirk Inzé Lab Frank Van Breusegem Lab Geert De Jaeger Lab Ive De Smet Lab Jenny Russinova Lab Lieven De Veylder Lab Mieke

    Original URL path: http://www.psb.vib-ugent.be/daniel-van-damme (2016-04-26)
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