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51. Opportunities for recombinant antigen and antibody expression in transgenic plants - technology assessment

Author

Schillberg, S., Twyman, R.M., Fischer, R., and Publica

Abstract

Plants are now gaining widespread acceptance as a general platform for the large-scale production of recombinant proteins. The principle has been demonstrated by the success of a diverse repertoire of proteins, with therapeutic molecules showing the most potential for added value. Over the past 10 years, several efficient plant-based expression systems have emerged. However, a number of issues remain to be addressed before plant bioreactors can be accepted and adopted widely in preference to the established microbial and mammalian platforms. Overcoming bottlenecks imposed by low yields, poor and inconsistent product quality and difficulties with downstream processing are the most important goals for researchers working in this field. The achievement of these goals is conditional on the development of extraction and processing steps that comply with GMP standards, including extensive quality assurance and control procedures. Such rigorous and validated standards should be combined with measures applied earlier in production to ensure product sustainability and quality, such as the use of master seed banking procedures. Moreover, there are several further challenges concerning topics of environmental impact, biosafety and risk assessment, which reflect the release of transgenic plants, as well the safety of the plant-derived products themselves. We are facing a growing demand for protein diagnostics and therapeutics, but lack the capacity to meet those demands using established facilities. A shift to plant bioreactors may, therefore, become necessary within the next few years, making it more imperative that the technical and regulatory limitations are addressed and solved. The production of pharmaceutical proteins in plants will only realize its huge potential if the products are provided at consistent high quality levels, allowing the delivery of clinical grade proteins that will gain regulatory approval and which can be used routinely in clinical trials.

Published
2005

53. Immunomodulation of polyamine biosynthesis in tobacco plants has a significant impact on polyamine levels and generates a dwarf phenotype

Author

Nölke, G., Schneider, B., Fischer, R., Schillberg, S., and Publica

Abstract

Ornithine decarboxylase (ODC) is a cytosolic enzyme that catalyses the direct decarboxylation of l-ornithine to putrescine, one of the rate-limiting steps of polyamine biosynthesis in plants. In the present study, an ODC-specific murine single-chain antibody fragment (scFvODC1) was generated by phage display technology. To evaluate the effect of the recombinant antibody fragment on ODC activity and polyamine levels, we produced transgenic tobacco plants that accumulated scFvODC1 in the cytosol. Expression levels of up to 4% total soluble protein (TSP) were achieved, resulting in the inhibition of up to 90% of endogenous ODC activity. A significant reduction in putrescine, spermidine and spermine levels was observed in transgenic lines producing high levels of scFvODC1. Furthermore, these lines showed developmental abnormalities and a dwarf phenotype. We show that the immunomodulation of enzyme activity is a powerful approach that can be used to alter complex and tightly controlled metabolic pathways, allowing specific steps in the pathway to be blocked and the resulting physiological effects to be investigated.

Published
2005

56. Molecular farming in plants: Host systems and expression technology

Author

Twyman, R.M., Stoger, E., Schillberg, S., Christou, P., Fischer, R., and Publica

Abstract

Plants provide an inexpensive and convenient system for the large-scale production of valuable recombinant proteins. This principle has been demonstrated by the commercial success of several first-generation products, and many others are currently under development. Over the past ten years, several efficient plant-based expression systems have emerged, and >100 recombinant proteins have now been produced in a range of different species. Plants have many advantages over other production systems, particularly in terms of practicality, economy and safety. However, several constraints that hinder the widespread use of plants as bioreactors remain to be addressed. Important factors include quality and homogeneity of the final product, the challenge of processing plant-derived pharmaceutical macromolecules under good manufacturing practice conditions and concerns about biosafety. Molecular farming in plants will only realize its huge potential if these constraints are removed through rigorous and detailed science-based studies.

Published
2003

57. Molecular Farming. Produktion hochwertiger Proteine in Pflanzen

Author

Mavituna, M., Schillberg, S., Fischer, R., and Publica

Abstract

Als Molecular Farming bezeichnet man die Produktion von hochwertigen Proteinen in Pflanzen. Neben Molecular Farming werden auch die Begriffe Gen-Farming, Bio-Pharming oder Molecular Pharming verwendet. Dazu wird ein Gen, das zuvor aus der Erbsubstanz anderer Spezies isoliert wurde, in das Genom einer Pflanze integriert. Diese genetisch veränderten, "tansgenen" Pflanzen produzieren dann aufgrund der Information des Fremdgens ein neues "rekombinantes" Protein und ermöglichen eine kostengünstige und sichere Proteinproduktion im großen Maßstab. Zur Produktion der rekombinanten Proteine werden intakte Pflanzen verwendet, die auf dem Feld oder im Gewächshaus angebaut werden, oder pflanzliche Zellkulturen, deren Kultivierung unter kontrollierten Bedingungen in Bioreaktoren erfolgt.

Published
2002

58. Antibody-based resistance to plant pathogens

Author

Schillberg, S., Zimmermann, S., Zhang, M.Y., Fischer, R., and Publica

Subjects
pathogen resistance, protein targeting, recombinant antibodies, transgenic plants
Abstract

Plant diseases are a major threat to the world food supply, as up to 15 per cent of production is lost to pathogens. In the past, disease control and the generation of resistant plant lines protected against viral, bacterial or fungal pathogens, was achieved using conventional breeding based on crossings, mutant screenings and backcrossing. Many approaches in this field have failed or the resistance obtained has been rapidly broken by the pathogens. Recent advances in molecular biotechnology have made it possible to obtain and to modify genes that are useful for generating disease resistant crops. Several strategies, including expression of pathogen-derived sequences or anti-pathogenic agents, have been developed to engineer improved pathogen resistance in transgenic plants. Antibody-based resistance is a novel strategy for generating transgenic plants resistant to pathogens. Decades ago it was shown that polyclonal and monoclonal antibodies can neutralize viruses, bacteria and selected fungi. This approach has been improved recently by the development of recombinant antibodies (rAbs). Crop resistance can be engineered by the expression of pathogen-specific antibodies, antibody fragments or antibody fusion proteins. The advantages of this approach are that rAbs can be engineered against almost any target molecule, and it has been demonstrated that expression of functional pathogen-specific rAbs in plants confers effective pathogen protection. The efficacy of antibody-based resistance was first shown for plant viruses and its application to other plant pathogens is becoming more established. However, successful use of antibodies to generate plant pathogen resistance relies on appropriate target selection, careful antibody design, efficient antibody expression, stability and targeting to appropriate cellular compartments.

Published
2001

60. Plasma membrane display of anti-viral single chain Fv fragments confers resistance to tobacco mosaic virus

Author

Schillberg, S., Zimmermann, S., Findley, K., Fischer, R., and Publica

Subjects
membrane anchor, tobacco mosaic virus, protein targeting, single chain antibody, transgenic plants
Abstract

We tested the hypothesis that membrane-anchored anti-viral antibodies can confer viral resistance to transgenic plants. A heterologous expression system was developed for plasma membrane targeting of anti-viral antibodies using mammalian transmembrane domains. A tobacco mosaic virus (TMV) neutralizing single-chain Fv antibody fragment (scFv24) was targeted to the endoplasmic reticulum and integrated into the plasma membrane of tobacco cells, using mammalian signal peptides and membrane receptor transmembrane domains. The human platelet-derived growth factor receptor (PDGFR) transmembrane domain or the T-cell receptor beta-domain (TcR beta) transmembrane domain was fused to the C-terminus of TMV-specific scFv24 to target expression of scFv24 as an extracellularly facing plasma membrane protein. Western blot and ELISA analyses were carried out to confirm functional expression of the recombinant fusion proteins scFv24-PDGFR and scFv24-TcR beta in transgenic tobacco suspension cultures and transgenic plants. Immunofluorescence and electron microscopy showed that the TcR beta transmembrane domain targeted scFv24 to the tobacco plasma membrane. Bioassays of viral infection showed that transgenic tobacco plants expressing scFv24-TcR beta were resistant to TMV infection. These results demonstrated that membrane anchored anti-viral antibody fragments are functional, can be targeted of the plasma membrane in planta and are a novel approach for engineering disease-resistant crops.

Published
2000

63. Apoplastic and cytosolic expression of full-size antibodies and antibody fragments in Nicotiana tabacum

Author

Schillberg, S., Zimmermann, S., Voss, A., Fischer, R., and Publica

Subjects
TMV, plant virus, Cytosolic expression, recombinant antibody, single chain antibody, transgenic plants
Abstract

We compared the expression of a functional recombinant TMV-specific full-size antibody (rAb29) in both the apoplast and cytosol of tobacco plants and a single chain antibody fragment (scFv29), derived from rAb29, was expressed in the cytosol. Cloned heavy and light chain cDNAs of full-size rAb29, which binds to coat protein monomers, were integrated into the plant expression vector pSS. The full-size rAb29 was expressed in the cytosol and targeted to the apoplast by including the original murine antibody leader sequences. Levels of functional full-size rAb29 expression were high in the apoplast (up to 8.5ng per gram leaf tissue), whereas cytosolic expression was low or at the ELISA detection limit. Sequences of the variable domains of rAb29 light and heavy chain were used to generate the single chain antibody scFv29, which was expressed in the periplasmic space of E. coli and showed the same binding specificity as full-size rAb29. In addition, scFv29 was functionally expressed in the cytosol of tobacco plants and plant derived scFv29 maintained same binding specificity to TMV-coat protein monomers as rAb29.

Published
1999

64. Expression and characterization of bispecific single chain Fv fragments produced in transgenic plants

Author

Fischer, R., Schumann, D., Zimmermann, S., Drossard, J., Sack, M., Schillberg, S., and Publica

Subjects
tobacco mosaic virus, protein targeting, recombinant antibodies, transgenic plants, bispecific single-chain antibody
Abstract

We describe the expression of the bispecific antibody biscFv2429 in transgenic suspension culture cells and tobacco plants. biscFv2429 consists of two single chain antibodies, scFv24 and scFv29, connected by the Trichoderma reesi cellobiohydrolase I linker. biscFv2429 binds two epitopes of tobacco mosaic virus (TMV): the scFv24 domain recognizes neotopes of intact virions, and the scFv29 domain recognizes a cryptotope of the TMV coat protein monomer. biscFv2429 was functionally expressed either in the cytosol (biscFv2429-cyt) or targeted to the apoplast using a murine leader peptide sequence (biscFv2429-apoplast). A third construct contained the C-terminal KDEL sequence for retention in the ER (biscFv2429-KDEL). Levels of cytoplasmic biscFv2429 expression levels were low. The highest levels of antibody expression were for apoplast targeted biscFv2429-apoplast and ER retained biscFv2429-KDEL, which reached a maximum expression level of 1.65% total soluble protein in transgenic plants. P lant expressed biscFv2429 retained both epitope specificities, and bispecificity and bivalency were confirmed by ELISA and surface plasmon resonance analysis. This study establishes plant cells as an expression system for bispecific single chain antibodies for use in medical and biological applications.

Published
1999

74. Contained molecular farming

Author

Anneli Ritala, Suvi Häkkinen, Kirsi-Marja Oksman-Caldentey, Raven, N., Schillberg, S., Henquet, M., and Bosch, D.

78. Towards molecular farming in the future: Moving from diagnostic protein and antibody production in microbes to plants

Author

Fischer, R., Drossard, J., Ulrich Commandeur, Schillberg, S., Emans, N., and Publica

Abstract

Molecular Farming of pharmaceuticals in plants has the potential to provide almost unlimited amounts of recombinant proteins for use in disease diagnosis and therapy. Transgenic plants are attracting interest as bioreactors for the inexpensive production of large amounts of safe, functional recombinant macromolecules, such as blood substitutes, vaccines and antibodies. In some cases, the function of recombinant proteins can be rapidly analysed by expression in microbes or by transient expression in intact or virally infected plants. Protein production can be increased by upscaling production in fermenters, using yeast or plant suspension cells or transient expression systems. Stable transgenic plants can be used to produce leaves or seeds rich in the recombinant protein for long term storage or direct processing. This demonstrates the promise for using plants as bioreactors for the molecular farming of recombinant therapeutics, diagnostics, blood substitutes and antibodies. We anticipa te that this technology has the potential to greatly benefit human health by making safe recombinant pharmaceuticals widely available.

81. Abstracts of presentations on plant protection issues at the xth international congress of virology: August 11-16,1996 Binyanei haOoma, Jerusalem, Israel Part 2 Plenary Lectures

Author

Ramirez, E., Hernádez, A., Ham, E., Barrón, B., Ding, S., Li, W., Shi, B., Symons, R., Li, Q., Ryu, K., Palukaitis, P., Kaplan, I., Gal-On, A., Kunhara, J., Otsubo, T., Tomaru, K., Tsuda, S., Kinta, M., Lin, T., Johnson, J., Porta, C., Taylor, K., Spall, V., Lomonossoff, G., Meiri, E., Raccah, B., Gaba, V., Ikegami, M., Kawashima, H., Murayama, A., Natsuaki, T., Kageyama, Y., Sugimura, N., Thole, V., Bol, J., Deiman, B., Koenen, A., Pleij, C., Chaleeprom, W., Bateson, M., Dale, J., Badge, J., Foster, G., Brunt, A., Robinson, D., Schubert, J., Rabenstein, F., Merits, A., Yeh, Shyi-Dong, Chiang, Chu-Hui, Wang, Ching-Hsien, Golshani, A., Ivanov, I., AbouHaidar, M., Rodoni, B., Harding, R., Oertel, U., Fuchs, E., Yang, S., Revere, F., Walter, J., Le Gall, O., Dunez, J., Candresse, T., Lot, H., Souche, S., Kasschau, Kristin, Carrington, James, Chiang, A., Turner, N., Hwang, D., Chachulska, A., Zagórski, W., Robaglia, C., Chrzanowska, M., Truve, E., Tamm, T., Saarma, M., Makinen, K., Wang, Aiming, Carrier, Karma, Wieczorek, Andrew, Huguenot, Claire, Sanfacon, Helene, Wurscher, M., Christensen, F., Prody, G., Sanchez-Navarro, J., Pallas, V., Carson, R., Dagless, E., Lock, S., Turner, R., Adams, M., Antoniw, J., Phillips, S., Kashiwazaki, S., Seal, S., Karamagioli, M., Druka, Amis, Makhdoom, Rahat, Yitang, Yan, Hull, Roger, Kikkert, M., Bodegom, P., Storms, M., van Lent, J., Kormelink, R., Goldbach, R., De Haan, P., De Rover, A., Van Schepen, A., Gielen, J., Van Grinsven, M., Livneh, O., Vardi, E., Kuznetsova, L., Aliskevicius, E., Stram, Y., Sela, I., Dinant, S., Kusiak, C., Blaise, F., Chupeau, Y., Astier, S., Albouy, J., Clifford, T., Taylor, S., Stratford, R., Xiao, X., Frenkel, M., Chu, P., Tabe, L., Shukla, D., Ward, C., Hwang, Duk-Ju, Turner, Nilgun, Mooney, A., MacFarlane, S., Twell, D., Jacquet, C., Ravelonandro, M., Bachelier, J., Almási, A., Böddi, B., Szigeti, Z., Ekes, M., Gaborianvi, R., Yankulova, Margarita, Ivanova, Lidia, Adam, G., Guelemerov, S., Nikolaeva, Velitchka, Atanassov, A., Prins, Marcel, de Haan, Peter, Goldbach, Rob, Duan, Y., Powell, C., Purcifull, D., Hiebert, E., Levine, B., Faktor, O., Zeituny, Silvy, Edelbaum, Orit, Sela, L., Marano, Maria-Rosa, Ratcliff, Frank, Baulcombe, David, van Oers, Monique, Linthorst, Huub, Bol, John, Schillberg, S., Zimmermann, S., Drossard, J., Schumann, D., Nahring, J., Fischer, R., Lapidot, Moshe, Arce-Johnson, Patricio, Rivera-Bustamante, Rafael, Beachy, Roger, Cassidy, Brandt, Flasinski, Stanislaw, Hajimorad, M., Wesley, Varsha, Angel-Diaz, J., Mayo, M., Hafner, G., May, G., Becker, D., Arntzen, C., Porter, J., Hellwald, K., Banerjee, N., Zaitlin, M., Wolf, D., Faure, J., Pilowsky, M., Cohen, S., Zelcer, A., Hardi, V., Tomassoli, L., Kaniewski, W., Mitsky, T., Barba, M., Hou, Y., Ursin, V., Sanders, R., Gilbertson, R., Gruden, Kristina, Štrukelj, Borut, Brzin, Joze, Ravnikar, Maja, Maganja, Darja, Kregar, Igor, Maki-Valkama, T., Pehu, T., Valkonen, J., Lento, K., Pehu, E., Lawson, E., Feldman, J., Zalewski, J., Kuittinen, T., Atiri, G., Romero, A., Arroyo, R., Soto, M., Martínez-Zapater, J., Ponz, F., Gilardi, P., López, L., García-Luque, I., Serra, M., Pang, Sheng-Zhi, Jan, Fuh-Jyh, Carney, Kim, Tricoli, David, Quemada, Hector, Gonsalves, Dennis, Aboul-Ata, A., Thouvenel, J., Marshall, D., Abo-El-Saad, Sh, Satour, M., Mazithulela, G., Mehlo, L., Gahakwa, D., Leech, M., Boulton, M., Davies, J., Christou, P., Huet, H., Sivamani, E., Ong, C., Chen, L., de Kochko, A., Beachy, R., Fauquet, C., Sithisarn-Burns, P., Maugeri, M., Smith, G., Handley, J., Graichen, K., Peterka, J., Chu, Paul, Larkin, Phil, Higgins, T., Wylie, S., Jones, M., Somsap, V., Loo, H., Li, D., Mathews, A., Dwyer, G., McCarthy, P., Hansen, J., Shiel, P., Zemetra, R., Wyatt, S., Berger, P., Ford, C., Collins, N., Rathjen, J., Shams-Bakhsh, M., Paltridge, N., Makkouk, K., El-Saied, M., El-Hariry, M., Salem, G., Soliman, N., Rishi, Narayan, Lodhi, G., Bishnoi, S., Sangwan, R., Sijen, Titia, Wellink, Joan, van Kammen, Ab, Scorza, R., Callahan, A., Levy, L., Polák, J., Oukropec, I., Kominek, P., Bitoova, M., Cardol, Erwin, Kormelink, Richard, Baulcombe, D., English, J., Davenport, G., Ruiz-Perez, T., Mueller, E., Nigul, L., Kelve, M., Fedorkin, O., Denisenko, O., Zelenina, D., Morozov, S., Atabekov, J., Laliberté, J., Wittmann, S., Plante, Daniel, Fortin, Marc, Chatel, H., Rodriguez-Alvarado, G., Garcia-Arenal, F., Roossinck, M., Gellatly, D., Rasochova, L., Aulik, M., Passmore, B., Falk, B., Miller, W., Lin, Na-Sheng, Lin, Biing-Yuan, Hsu, Yau-Heiu, Tamada, T., Kiguchi, T., Saito, M., Kusume, T., Uchino, H., Taliansky, M., DiSerio, F., Daròs, J., Ragozzino, A., Floras, R., Hohn, Thomas, Chen, Gang, Rothnie, Helen, Corsten, Sandra, Fütterer, Johannes, Mikhailov, M., Bishop, D., Roy, P., Ramirez, E., Hernádez, A., Ham, E., Barrón, B., Ding, S., Li, W., Shi, B., Symons, R., Li, Q., Ryu, K., Palukaitis, P., Kaplan, I., Gal-On, A., Kunhara, J., Otsubo, T., Tomaru, K., Tsuda, S., Kinta, M., Lin, T., Johnson, J., Porta, C., Taylor, K., Spall, V., Lomonossoff, G., Meiri, E., Raccah, B., Gaba, V., Ikegami, M., Kawashima, H., Murayama, A., Natsuaki, T., Kageyama, Y., Sugimura, N., Thole, V., Bol, J., Deiman, B., Koenen, A., Pleij, C., Chaleeprom, W., Bateson, M., Dale, J., Badge, J., Foster, G., Brunt, A., Robinson, D., Schubert, J., Rabenstein, F., Merits, A., Yeh, Shyi-Dong, Chiang, Chu-Hui, Wang, Ching-Hsien, Golshani, A., Ivanov, I., AbouHaidar, M., Rodoni, B., Harding, R., Oertel, U., Fuchs, E., Yang, S., Revere, F., Walter, J., Le Gall, O., Dunez, J., Candresse, T., Lot, H., Souche, S., Kasschau, Kristin, Carrington, James, Chiang, A., Turner, N., Hwang, D., Chachulska, A., Zagórski, W., Robaglia, C., Chrzanowska, M., Truve, E., Tamm, T., Saarma, M., Makinen, K., Wang, Aiming, Carrier, Karma, Wieczorek, Andrew, Huguenot, Claire, Sanfacon, Helene, Wurscher, M., Christensen, F., Prody, G., Sanchez-Navarro, J., Pallas, V., Carson, R., Dagless, E., Lock, S., Turner, R., Adams, M., Antoniw, J., Phillips, S., Kashiwazaki, S., Seal, S., Karamagioli, M., Druka, Amis, Makhdoom, Rahat, Yitang, Yan, Hull, Roger, Kikkert, M., Bodegom, P., Storms, M., van Lent, J., Kormelink, R., Goldbach, R., De Haan, P., De Rover, A., Van Schepen, A., Gielen, J., Van Grinsven, M., Livneh, O., Vardi, E., Kuznetsova, L., Aliskevicius, E., Stram, Y., Sela, I., Dinant, S., Kusiak, C., Blaise, F., Chupeau, Y., Astier, S., Albouy, J., Clifford, T., Taylor, S., Stratford, R., Xiao, X., Frenkel, M., Chu, P., Tabe, L., Shukla, D., Ward, C., Hwang, Duk-Ju, Turner, Nilgun, Mooney, A., MacFarlane, S., Twell, D., Jacquet, C., Ravelonandro, M., Bachelier, J., Almási, A., Böddi, B., Szigeti, Z., Ekes, M., Gaborianvi, R., Yankulova, Margarita, Ivanova, Lidia, Adam, G., Guelemerov, S., Nikolaeva, Velitchka, Atanassov, A., Prins, Marcel, de Haan, Peter, Goldbach, Rob, Duan, Y., Powell, C., Purcifull, D., Hiebert, E., Levine, B., Faktor, O., Zeituny, Silvy, Edelbaum, Orit, Sela, L., Marano, Maria-Rosa, Ratcliff, Frank, Baulcombe, David, van Oers, Monique, Linthorst, Huub, Bol, John, Schillberg, S., Zimmermann, S., Drossard, J., Schumann, D., Nahring, J., Fischer, R., Lapidot, Moshe, Arce-Johnson, Patricio, Rivera-Bustamante, Rafael, Beachy, Roger, Cassidy, Brandt, Flasinski, Stanislaw, Hajimorad, M., Wesley, Varsha, Angel-Diaz, J., Mayo, M., Hafner, G., May, G., Becker, D., Arntzen, C., Porter, J., Hellwald, K., Banerjee, N., Zaitlin, M., Wolf, D., Faure, J., Pilowsky, M., Cohen, S., Zelcer, A., Hardi, V., Tomassoli, L., Kaniewski, W., Mitsky, T., Barba, M., Hou, Y., Ursin, V., Sanders, R., Gilbertson, R., Gruden, Kristina, Štrukelj, Borut, Brzin, Joze, Ravnikar, Maja, Maganja, Darja, Kregar, Igor, Maki-Valkama, T., Pehu, T., Valkonen, J., Lento, K., Pehu, E., Lawson, E., Feldman, J., Zalewski, J., Kuittinen, T., Atiri, G., Romero, A., Arroyo, R., Soto, M., Martínez-Zapater, J., Ponz, F., Gilardi, P., López, L., García-Luque, I., Serra, M., Pang, Sheng-Zhi, Jan, Fuh-Jyh, Carney, Kim, Tricoli, David, Quemada, Hector, Gonsalves, Dennis, Aboul-Ata, A., Thouvenel, J., Marshall, D., Abo-El-Saad, Sh, Satour, M., Mazithulela, G., Mehlo, L., Gahakwa, D., Leech, M., Boulton, M., Davies, J., Christou, P., Huet, H., Sivamani, E., Ong, C., Chen, L., de Kochko, A., Beachy, R., Fauquet, C., Sithisarn-Burns, P., Maugeri, M., Smith, G., Handley, J., Graichen, K., Peterka, J., Chu, Paul, Larkin, Phil, Higgins, T., Wylie, S., Jones, M., Somsap, V., Loo, H., Li, D., Mathews, A., Dwyer, G., McCarthy, P., Hansen, J., Shiel, P., Zemetra, R., Wyatt, S., Berger, P., Ford, C., Collins, N., Rathjen, J., Shams-Bakhsh, M., Paltridge, N., Makkouk, K., El-Saied, M., El-Hariry, M., Salem, G., Soliman, N., Rishi, Narayan, Lodhi, G., Bishnoi, S., Sangwan, R., Sijen, Titia, Wellink, Joan, van Kammen, Ab, Scorza, R., Callahan, A., Levy, L., Polák, J., Oukropec, I., Kominek, P., Bitoova, M., Cardol, Erwin, Kormelink, Richard, Baulcombe, D., English, J., Davenport, G., Ruiz-Perez, T., Mueller, E., Nigul, L., Kelve, M., Fedorkin, O., Denisenko, O., Zelenina, D., Morozov, S., Atabekov, J., Laliberté, J., Wittmann, S., Plante, Daniel, Fortin, Marc, Chatel, H., Rodriguez-Alvarado, G., Garcia-Arenal, F., Roossinck, M., Gellatly, D., Rasochova, L., Aulik, M., Passmore, B., Falk, B., Miller, W., Lin, Na-Sheng, Lin, Biing-Yuan, Hsu, Yau-Heiu, Tamada, T., Kiguchi, T., Saito, M., Kusume, T., Uchino, H., Taliansky, M., DiSerio, F., Daròs, J., Ragozzino, A., Floras, R., Hohn, Thomas, Chen, Gang, Rothnie, Helen, Corsten, Sandra, Fütterer, Johannes, Mikhailov, M., Bishop, D., and Roy, P.

83. Development of an optimized tetracycline-inducible expression system to increase the accumulation of interleukin-10 in tobacco BY-2 suspension cells

Author

Bortesi Luisa, Rademacher Thomas, Schiermeyer Andreas, Schuster Flora, Pezzotti Mario, and Schillberg Stefan

Subjects
Biotechnology, TP248.13-248.65
Abstract

Abstract Background Plant cell suspension cultures can be used for the production of valuable pharmaceutical and industrial proteins. When the recombinant protein is secreted into the culture medium, restricting expression to a defined growth phase can improve both the quality and quantity of the recovered product by minimizing proteolytic activity. Temporal restriction is also useful for recombinant proteins whose constitutive expression affects cell growth and viability, such as viral interleukin-10 (vIL-10). Results We have developed a novel, tetracycline-inducible system suitable for tobacco BY-2 suspension cells which increases the yields of vIL-10. The new system is based on a binary vector that is easier to handle than conventional vectors, contains an enhanced inducible promoter and 5′-UTR to improve yields, and incorporates a constitutively-expressed visible marker gene to allow the rapid and straightforward selection of the most promising transformed clones. Stable transformation of BY-2 cells with this vector, without extensive optimization of the induction conditions, led to a 3.5 fold increase in vIL-10 levels compared to constitutive expression in the same host. Conclusions We have developed an effective and straightforward molecular farming platform technology that improves both the quality and the quantity of recombinant proteins produced in plant cells, particularly those whose constitutive expression has a negative impact on plant growth and development. Although we tested the platform using vIL-10 produced in BY-2 cells, it can be applied to other host/product combinations and is also useful for basic research requiring strictly controlled transgene expression.

Published
2012
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84. A membrane-bound matrix-metalloproteinase from Nicotiana tabacum cv. BY-2 is induced by bacterial pathogens

Author

Wahner Verena, Otte Burkhard, Mandal Manoj K, Hartenstein Hanna, Schiermeyer Andreas, and Schillberg Stefan

Subjects
Botany, QK1-989
Abstract

Abstract Background Plant matrix metalloproteinases (MMP) are conserved proteolytic enzymes found in a wide range of monocotyledonous and dicotyledonous plant species. Acting on the plant extracellular matrix, they play crucial roles in many aspects of plant physiology including growth, development and the response to stresses such as pathogen attack. Results We have identified the first tobacco MMP, designated NtMMP1, and have isolated the corresponding cDNA sequence from the tobacco suspension cell line BY-2. The overall domain structure of NtMMP1 is similar to known MMP sequences, although certain features suggest it may be constitutively active rather than dependent on proteolytic processing. The protein appears to be expressed in two forms with different molecular masses, both of which are enzymatically active as determined by casein zymography. Exchanging the catalytic domain of NtMMP1 with green fluorescent protein (GFP) facilitated subcellular localization by confocal laser scanning microscopy, showing the protein is normally inserted into the plasma membrane. The NtMMP1 gene is expressed constitutively at a low level but can be induced by exposure to bacterial pathogens. Conclusion Our biochemical analysis of NtMMP1 together with bioinformatic data on the primary sequence indicate that NtMMP1 is a constitutively-active protease. Given its induction in response to bacterial pathogens and its localization in the plasma membrane, we propose a role in pathogen defense at the cell periphery.

Published
2009
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85. Viral and murine interleukin-10 are correctly processed and retain their biological activity when produced in tobacco

Author

Avesani Linda, Stadlmann Johannes, Raven Nicole, Schuster Flora, Rossato Marzia, Bortesi Luisa, Falorni Alberto, Bazzoni Flavia, Bock Ralph, Schillberg Stefan, and Pezzotti Mario

Subjects
Biotechnology, TP248.13-248.65
Abstract

Abstract Background Interleukin-10 (IL-10) is a potent anti-inflammatory cytokine, with therapeutic applications in several autoimmune and inflammatory diseases. Oral administration of this cytokine alone, or in combination with disease-associated autoantigens could confer protection form the onset of a specific autoimmune disease through the induction of oral tolerance. Transgenic plants are attractive systems for production of therapeutic proteins because of the ability to do large scale-up at low cost, and the low maintenance requirements. They are highly amenable to oral administration and could become effective delivery systems without extensive protein purification. We investigated the ability of tobacco plants to produce high levels of biologically-active viral and murine IL-10. Results Three different subcellular targeting strategies were assessed in transient expression experiments, and stable transgenic tobacco plants were generated with the constructs that yielded the highest accumulation levels by targeting the recombinant proteins to the endoplasmic reticulum. The best yields using this strategy in T1 plants were 10.8 and 37.0 μg/g fresh leaf weight for viral and murine IL-10, respectively. The recombinant proteins were purified from transgenic leaf material and characterized in terms of their N-glycan composition, dimerization and biological activity in in vitro assays. Both molecules formed stable dimers, were able to activate the IL-10 signaling pathway and to induce specific anti-inflammatory responses in mouse J774 macrophage cells. Conclusion Tobacco plants are able to correctly process viral and murine IL-10 into biologically active dimers, therefore representing a suitable platform for the production for these cytokines. The accumulation levels obtained are high enough to allow delivery of an immunologically relevant dose of IL-10 in a reasonable amount of leaf material, without extensive purification. This study paves the way to performing feeding studies in mouse models of autoimmune diseases, that will allow the evaluation the immunomodulatory properties and effectiveness of the viral IL-10 in inducing oral tolerance compared to the murine protein.

Published
2009
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86. Advances in the Chemical Synthesis of Carbohydrates and Glycoconjugates

Author

Malik, Ankita, Seeberger, Peter H., Varón Silva, Daniel, Scheper, Thomas, Series Editor, Belkin, Shimshon, Editorial Board Member, Bley, Thomas, Editorial Board Member, Bohlmann, Jörg, Editorial Board Member, Gu, Man Bock, Editorial Board Member, Hu, Wei-Shou, Editorial Board Member, Mattiasson, Bo, Editorial Board Member, Olsson, Lisbeth, Editorial Board Member, Seitz, Harald, Editorial Board Member, Silva, Ana Catarina, Editorial Board Member, Ulber, Roland, Editorial Board Member, Zeng, An-Ping, Editorial Board Member, Zhong, Jian-Jiang, Editorial Board Member, Zhou, Weichang, Editorial Board Member, Rapp, Erdmann, editor, Reichl, Udo, editor, Alagesan, K., With Contri.By, Betenbaugh, M. J., With Contri.By, Butler, M., With Contri.By, Buyel, J. F., With Contri.By, Cajic, S., With Contri.By, Callewaert, N., With Contri.By, Can, Ö., With Contri.By, Cox, M. M. J., With Contri.By, DeLisa, M. P., With Contri.By, De Wachter, C., With Contri.By, Elling, L., With Contri.By, Fischer, R., With Contri.By, Flitsch, S. L., With Contri.By, Frohnmeyer, H., With Contri.By, Gottschalk, J., With Contri.By, Goyette-Desjardins, G., With Contri.By, Heffner, K. M., With Contri.By, Hizal, D. B., With Contri.By, Hoffmann, M., With Contri.By, Holland, T., With Contri.By, Jefferis, R., With Contri.By, Kolarich, D., With Contri.By, Laaf, D., With Contri.By, Lepenies, B., With Contri.By, Malik, A., With Contri.By, Martinez, J. E. R., With Contri.By, Mills, D. C., With Contri.By, Moeller, T. D., With Contri.By, Palomares, L. A., With Contri.By, Pralow, A., With Contri.By, RamÚrez, O. T., With Contri.By, Rapp, E., With Contri.By, Reichl, U., With Contri.By, Rexer, T., With Contri.By, Sack, M., With Contri.By, Schillberg, S., With Contri.By, Schön, K., With Contri.By, Seeberger, P. H., With Contri.By, Srivastava, I. K., With Contri.By, Stoger, E., With Contri.By, Thomas, B., With Contri.By, Twyman, R. M., With Contri.By, Van Landuyt, L., With Contri.By, Varón Silva, D., With Contri.By, Wang, Q., With Contri.By, Weyant, K. B., With Contri.By, and Yates, L. E., With Contri.By

Published
2021
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87. Enzymatic Synthesis of Glycans and Glycoconjugates

Author

Rexer, Thomas, Laaf, Dominic, Gottschalk, Johannes, Frohnmeyer, Hannes, Rapp, Erdmann, Elling, Lothar, Scheper, Thomas, Series Editor, Belkin, Shimshon, Editorial Board Member, Bley, Thomas, Editorial Board Member, Bohlmann, Jörg, Editorial Board Member, Gu, Man Bock, Editorial Board Member, Hu, Wei-Shou, Editorial Board Member, Mattiasson, Bo, Editorial Board Member, Olsson, Lisbeth, Editorial Board Member, Seitz, Harald, Editorial Board Member, Silva, Ana Catarina, Editorial Board Member, Ulber, Roland, Editorial Board Member, Zeng, An-Ping, Editorial Board Member, Zhong, Jian-Jiang, Editorial Board Member, Zhou, Weichang, Editorial Board Member, Rapp, Erdmann, editor, Reichl, Udo, editor, Alagesan, K., With Contri.By, Betenbaugh, M. J., With Contri.By, Butler, M., With Contri.By, Buyel, J. F., With Contri.By, Cajic, S., With Contri.By, Callewaert, N., With Contri.By, Can, Ö., With Contri.By, Cox, M. M. J., With Contri.By, DeLisa, M. P., With Contri.By, De Wachter, C., With Contri.By, Elling, L., With Contri.By, Fischer, R., With Contri.By, Flitsch, S. L., With Contri.By, Frohnmeyer, H., With Contri.By, Gottschalk, J., With Contri.By, Goyette-Desjardins, G., With Contri.By, Heffner, K. M., With Contri.By, Hizal, D. B., With Contri.By, Hoffmann, M., With Contri.By, Holland, T., With Contri.By, Jefferis, R., With Contri.By, Kolarich, D., With Contri.By, Laaf, D., With Contri.By, Lepenies, B., With Contri.By, Malik, A., With Contri.By, Martinez, J. E. R., With Contri.By, Mills, D. C., With Contri.By, Moeller, T. D., With Contri.By, Palomares, L. A., With Contri.By, Pralow, A., With Contri.By, RamÚrez, O. T., With Contri.By, Rapp, E., With Contri.By, Reichl, U., With Contri.By, Rexer, T., With Contri.By, Sack, M., With Contri.By, Schillberg, S., With Contri.By, Schön, K., With Contri.By, Seeberger, P. H., With Contri.By, Srivastava, I. K., With Contri.By, Stoger, E., With Contri.By, Thomas, B., With Contri.By, Twyman, R. M., With Contri.By, Van Landuyt, L., With Contri.By, Varón Silva, D., With Contri.By, Wang, Q., With Contri.By, Weyant, K. B., With Contri.By, and Yates, L. E., With Contri.By

Published
2021
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88. Interplay of Carbohydrate and Carrier in Antibacterial Glycoconjugate Vaccines

Author

Moeller, Tyler D., Weyant, Kevin B., DeLisa, Matthew P., Scheper, Thomas, Series Editor, Belkin, Shimshon, Editorial Board Member, Bley, Thomas, Editorial Board Member, Bohlmann, Jörg, Editorial Board Member, Gu, Man Bock, Editorial Board Member, Hu, Wei-Shou, Editorial Board Member, Mattiasson, Bo, Editorial Board Member, Olsson, Lisbeth, Editorial Board Member, Seitz, Harald, Editorial Board Member, Silva, Ana Catarina, Editorial Board Member, Ulber, Roland, Editorial Board Member, Zeng, An-Ping, Editorial Board Member, Zhong, Jian-Jiang, Editorial Board Member, Zhou, Weichang, Editorial Board Member, Rapp, Erdmann, editor, Reichl, Udo, editor, Alagesan, K., With Contri.By, Betenbaugh, M. J., With Contri.By, Butler, M., With Contri.By, Buyel, J. F., With Contri.By, Cajic, S., With Contri.By, Callewaert, N., With Contri.By, Can, Ö., With Contri.By, Cox, M. M. J., With Contri.By, DeLisa, M. P., With Contri.By, De Wachter, C., With Contri.By, Elling, L., With Contri.By, Fischer, R., With Contri.By, Flitsch, S. L., With Contri.By, Frohnmeyer, H., With Contri.By, Gottschalk, J., With Contri.By, Goyette-Desjardins, G., With Contri.By, Heffner, K. M., With Contri.By, Hizal, D. B., With Contri.By, Hoffmann, M., With Contri.By, Holland, T., With Contri.By, Jefferis, R., With Contri.By, Kolarich, D., With Contri.By, Laaf, D., With Contri.By, Lepenies, B., With Contri.By, Malik, A., With Contri.By, Martinez, J. E. R., With Contri.By, Mills, D. C., With Contri.By, Moeller, T. D., With Contri.By, Palomares, L. A., With Contri.By, Pralow, A., With Contri.By, RamÚrez, O. T., With Contri.By, Rapp, E., With Contri.By, Reichl, U., With Contri.By, Rexer, T., With Contri.By, Sack, M., With Contri.By, Schillberg, S., With Contri.By, Schön, K., With Contri.By, Seeberger, P. H., With Contri.By, Srivastava, I. K., With Contri.By, Stoger, E., With Contri.By, Thomas, B., With Contri.By, Twyman, R. M., With Contri.By, Van Landuyt, L., With Contri.By, Varón Silva, D., With Contri.By, Wang, Q., With Contri.By, Weyant, K. B., With Contri.By, and Yates, L. E., With Contri.By

Published
2021
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89. Glyco-Engineering of Plant-Based Expression Systems

Author

Fischer, Rainer, Holland, Tanja, Sack, Markus, Schillberg, Stefan, Stoger, Eva, Twyman, Richard M., Buyel, Johannes F., Scheper, Thomas, Series Editor, Belkin, Shimshon, Editorial Board Member, Bley, Thomas, Editorial Board Member, Bohlmann, Jörg, Editorial Board Member, Gu, Man Bock, Editorial Board Member, Hu, Wei-Shou, Editorial Board Member, Mattiasson, Bo, Editorial Board Member, Olsson, Lisbeth, Editorial Board Member, Seitz, Harald, Editorial Board Member, Silva, Ana Catarina, Editorial Board Member, Ulber, Roland, Editorial Board Member, Zeng, An-Ping, Editorial Board Member, Zhong, Jian-Jiang, Editorial Board Member, Zhou, Weichang, Editorial Board Member, Rapp, Erdmann, editor, Reichl, Udo, editor, Alagesan, K., With Contri.By, Betenbaugh, M. J., With Contri.By, Butler, M., With Contri.By, Buyel, J. F., With Contri.By, Cajic, S., With Contri.By, Callewaert, N., With Contri.By, Can, Ö., With Contri.By, Cox, M. M. J., With Contri.By, DeLisa, M. P., With Contri.By, De Wachter, C., With Contri.By, Elling, L., With Contri.By, Fischer, R., With Contri.By, Flitsch, S. L., With Contri.By, Frohnmeyer, H., With Contri.By, Gottschalk, J., With Contri.By, Goyette-Desjardins, G., With Contri.By, Heffner, K. M., With Contri.By, Hizal, D. B., With Contri.By, Hoffmann, M., With Contri.By, Holland, T., With Contri.By, Jefferis, R., With Contri.By, Kolarich, D., With Contri.By, Laaf, D., With Contri.By, Lepenies, B., With Contri.By, Malik, A., With Contri.By, Martinez, J. E. R., With Contri.By, Mills, D. C., With Contri.By, Moeller, T. D., With Contri.By, Palomares, L. A., With Contri.By, Pralow, A., With Contri.By, RamÚrez, O. T., With Contri.By, Rapp, E., With Contri.By, Reichl, U., With Contri.By, Rexer, T., With Contri.By, Sack, M., With Contri.By, Schillberg, S., With Contri.By, Schön, K., With Contri.By, Seeberger, P. H., With Contri.By, Srivastava, I. K., With Contri.By, Stoger, E., With Contri.By, Thomas, B., With Contri.By, Twyman, R. M., With Contri.By, Van Landuyt, L., With Contri.By, Varón Silva, D., With Contri.By, Wang, Q., With Contri.By, Weyant, K. B., With Contri.By, and Yates, L. E., With Contri.By

Published
2021
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90. Engineering of Yeast Glycoprotein Expression

Author

De Wachter, Charlot, Van Landuyt, Linde, Callewaert, Nico, Scheper, Thomas, Series Editor, Belkin, Shimshon, Editorial Board Member, Bley, Thomas, Editorial Board Member, Bohlmann, Jörg, Editorial Board Member, Gu, Man Bock, Editorial Board Member, Hu, Wei-Shou, Editorial Board Member, Mattiasson, Bo, Editorial Board Member, Olsson, Lisbeth, Editorial Board Member, Seitz, Harald, Editorial Board Member, Silva, Ana Catarina, Editorial Board Member, Ulber, Roland, Editorial Board Member, Zeng, An-Ping, Editorial Board Member, Zhong, Jian-Jiang, Editorial Board Member, Zhou, Weichang, Editorial Board Member, Rapp, Erdmann, editor, Reichl, Udo, editor, Alagesan, K., With Contri.By, Betenbaugh, M. J., With Contri.By, Butler, M., With Contri.By, Buyel, J. F., With Contri.By, Cajic, S., With Contri.By, Callewaert, N., With Contri.By, Can, Ö., With Contri.By, Cox, M. M. J., With Contri.By, DeLisa, M. P., With Contri.By, De Wachter, C., With Contri.By, Elling, L., With Contri.By, Fischer, R., With Contri.By, Flitsch, S. L., With Contri.By, Frohnmeyer, H., With Contri.By, Gottschalk, J., With Contri.By, Goyette-Desjardins, G., With Contri.By, Heffner, K. M., With Contri.By, Hizal, D. B., With Contri.By, Hoffmann, M., With Contri.By, Holland, T., With Contri.By, Jefferis, R., With Contri.By, Kolarich, D., With Contri.By, Laaf, D., With Contri.By, Lepenies, B., With Contri.By, Malik, A., With Contri.By, Martinez, J. E. R., With Contri.By, Mills, D. C., With Contri.By, Moeller, T. D., With Contri.By, Palomares, L. A., With Contri.By, Pralow, A., With Contri.By, RamÚrez, O. T., With Contri.By, Rapp, E., With Contri.By, Reichl, U., With Contri.By, Rexer, T., With Contri.By, Sack, M., With Contri.By, Schillberg, S., With Contri.By, Schön, K., With Contri.By, Seeberger, P. H., With Contri.By, Srivastava, I. K., With Contri.By, Stoger, E., With Contri.By, Thomas, B., With Contri.By, Twyman, R. M., With Contri.By, Van Landuyt, L., With Contri.By, Varón Silva, D., With Contri.By, Wang, Q., With Contri.By, Weyant, K. B., With Contri.By, and Yates, L. E., With Contri.By

Published
2021
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91. Bacterial Glycoengineering as a Biosynthetic Route to Customized Glycomolecules

Author

Yates, Laura E., Mills, Dominic C., DeLisa, Matthew P., Scheper, Thomas, Series Editor, Belkin, Shimshon, Editorial Board Member, Bley, Thomas, Editorial Board Member, Bohlmann, Jörg, Editorial Board Member, Gu, Man Bock, Editorial Board Member, Hu, Wei-Shou, Editorial Board Member, Mattiasson, Bo, Editorial Board Member, Olsson, Lisbeth, Editorial Board Member, Seitz, Harald, Editorial Board Member, Silva, Ana Catarina, Editorial Board Member, Ulber, Roland, Editorial Board Member, Zeng, An-Ping, Editorial Board Member, Zhong, Jian-Jiang, Editorial Board Member, Zhou, Weichang, Editorial Board Member, Rapp, Erdmann, editor, Reichl, Udo, editor, Alagesan, K., With Contri.By, Betenbaugh, M. J., With Contri.By, Butler, M., With Contri.By, Buyel, J. F., With Contri.By, Cajic, S., With Contri.By, Callewaert, N., With Contri.By, Can, Ö., With Contri.By, Cox, M. M. J., With Contri.By, DeLisa, M. P., With Contri.By, De Wachter, C., With Contri.By, Elling, L., With Contri.By, Fischer, R., With Contri.By, Flitsch, S. L., With Contri.By, Frohnmeyer, H., With Contri.By, Gottschalk, J., With Contri.By, Goyette-Desjardins, G., With Contri.By, Heffner, K. M., With Contri.By, Hizal, D. B., With Contri.By, Hoffmann, M., With Contri.By, Holland, T., With Contri.By, Jefferis, R., With Contri.By, Kolarich, D., With Contri.By, Laaf, D., With Contri.By, Lepenies, B., With Contri.By, Malik, A., With Contri.By, Martinez, J. E. R., With Contri.By, Mills, D. C., With Contri.By, Moeller, T. D., With Contri.By, Palomares, L. A., With Contri.By, Pralow, A., With Contri.By, RamÚrez, O. T., With Contri.By, Rapp, E., With Contri.By, Reichl, U., With Contri.By, Rexer, T., With Contri.By, Sack, M., With Contri.By, Schillberg, S., With Contri.By, Schön, K., With Contri.By, Seeberger, P. H., With Contri.By, Srivastava, I. K., With Contri.By, Stoger, E., With Contri.By, Thomas, B., With Contri.By, Twyman, R. M., With Contri.By, Van Landuyt, L., With Contri.By, Varón Silva, D., With Contri.By, Wang, Q., With Contri.By, Weyant, K. B., With Contri.By, and Yates, L. E., With Contri.By

Published
2021
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92. Recombinant Proteins and Monoclonal Antibodies

Author

Jefferis, Roy, Scheper, Thomas, Series Editor, Belkin, Shimshon, Editorial Board Member, Bley, Thomas, Editorial Board Member, Bohlmann, Jörg, Editorial Board Member, Gu, Man Bock, Editorial Board Member, Hu, Wei-Shou, Editorial Board Member, Mattiasson, Bo, Editorial Board Member, Olsson, Lisbeth, Editorial Board Member, Seitz, Harald, Editorial Board Member, Silva, Ana Catarina, Editorial Board Member, Ulber, Roland, Editorial Board Member, Zeng, An-Ping, Editorial Board Member, Zhong, Jian-Jiang, Editorial Board Member, Zhou, Weichang, Editorial Board Member, Rapp, Erdmann, editor, Reichl, Udo, editor, Alagesan, K., With Contri.By, Betenbaugh, M. J., With Contri.By, Butler, M., With Contri.By, Buyel, J. F., With Contri.By, Cajic, S., With Contri.By, Callewaert, N., With Contri.By, Can, Ö., With Contri.By, Cox, M. M. J., With Contri.By, DeLisa, M. P., With Contri.By, De Wachter, C., With Contri.By, Elling, L., With Contri.By, Fischer, R., With Contri.By, Flitsch, S. L., With Contri.By, Frohnmeyer, H., With Contri.By, Gottschalk, J., With Contri.By, Goyette-Desjardins, G., With Contri.By, Heffner, K. M., With Contri.By, Hizal, D. B., With Contri.By, Hoffmann, M., With Contri.By, Holland, T., With Contri.By, Jefferis, R., With Contri.By, Kolarich, D., With Contri.By, Laaf, D., With Contri.By, Lepenies, B., With Contri.By, Malik, A., With Contri.By, Martinez, J. E. R., With Contri.By, Mills, D. C., With Contri.By, Moeller, T. D., With Contri.By, Palomares, L. A., With Contri.By, Pralow, A., With Contri.By, RamÚrez, O. T., With Contri.By, Rapp, E., With Contri.By, Reichl, U., With Contri.By, Rexer, T., With Contri.By, Sack, M., With Contri.By, Schillberg, S., With Contri.By, Schön, K., With Contri.By, Seeberger, P. H., With Contri.By, Srivastava, I. K., With Contri.By, Stoger, E., With Contri.By, Thomas, B., With Contri.By, Twyman, R. M., With Contri.By, Van Landuyt, L., With Contri.By, Varón Silva, D., With Contri.By, Wang, Q., With Contri.By, Weyant, K. B., With Contri.By, and Yates, L. E., With Contri.By

Published
2021
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93. Glycobiotechnology of the Insect Cell-Baculovirus Expression System Technology

Author

Palomares, Laura A., Srivastava, Indresh K., Ramírez, Octavio T., Cox, Manon M. J., Scheper, Thomas, Series Editor, Belkin, Shimshon, Editorial Board Member, Bley, Thomas, Editorial Board Member, Bohlmann, Jörg, Editorial Board Member, Gu, Man Bock, Editorial Board Member, Hu, Wei-Shou, Editorial Board Member, Mattiasson, Bo, Editorial Board Member, Olsson, Lisbeth, Editorial Board Member, Seitz, Harald, Editorial Board Member, Silva, Ana Catarina, Editorial Board Member, Ulber, Roland, Editorial Board Member, Zeng, An-Ping, Editorial Board Member, Zhong, Jian-Jiang, Editorial Board Member, Zhou, Weichang, Editorial Board Member, Rapp, Erdmann, editor, Reichl, Udo, editor, Alagesan, K., With Contri.By, Betenbaugh, M. J., With Contri.By, Butler, M., With Contri.By, Buyel, J. F., With Contri.By, Cajic, S., With Contri.By, Callewaert, N., With Contri.By, Can, Ö., With Contri.By, Cox, M. M. J., With Contri.By, DeLisa, M. P., With Contri.By, De Wachter, C., With Contri.By, Elling, L., With Contri.By, Fischer, R., With Contri.By, Flitsch, S. L., With Contri.By, Frohnmeyer, H., With Contri.By, Gottschalk, J., With Contri.By, Goyette-Desjardins, G., With Contri.By, Heffner, K. M., With Contri.By, Hizal, D. B., With Contri.By, Hoffmann, M., With Contri.By, Holland, T., With Contri.By, Jefferis, R., With Contri.By, Kolarich, D., With Contri.By, Laaf, D., With Contri.By, Lepenies, B., With Contri.By, Malik, A., With Contri.By, Martinez, J. E. R., With Contri.By, Mills, D. C., With Contri.By, Moeller, T. D., With Contri.By, Palomares, L. A., With Contri.By, Pralow, A., With Contri.By, RamÚrez, O. T., With Contri.By, Rapp, E., With Contri.By, Reichl, U., With Contri.By, Rexer, T., With Contri.By, Sack, M., With Contri.By, Schillberg, S., With Contri.By, Schön, K., With Contri.By, Seeberger, P. H., With Contri.By, Srivastava, I. K., With Contri.By, Stoger, E., With Contri.By, Thomas, B., With Contri.By, Twyman, R. M., With Contri.By, Van Landuyt, L., With Contri.By, Varón Silva, D., With Contri.By, Wang, Q., With Contri.By, Weyant, K. B., With Contri.By, and Yates, L. E., With Contri.By

Published
2021
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94. Animal Cell Expression Systems

Author

Butler, M., Reichl, U., Scheper, Thomas, Series Editor, Belkin, Shimshon, Editorial Board Member, Bley, Thomas, Editorial Board Member, Bohlmann, Jörg, Editorial Board Member, Gu, Man Bock, Editorial Board Member, Hu, Wei-Shou, Editorial Board Member, Mattiasson, Bo, Editorial Board Member, Olsson, Lisbeth, Editorial Board Member, Seitz, Harald, Editorial Board Member, Silva, Ana Catarina, Editorial Board Member, Ulber, Roland, Editorial Board Member, Zeng, An-Ping, Editorial Board Member, Zhong, Jian-Jiang, Editorial Board Member, Zhou, Weichang, Editorial Board Member, Rapp, Erdmann, editor, Reichl, Udo, editor, Alagesan, K., With Contri.By, Betenbaugh, M. J., With Contri.By, Butler, M., With Contri.By, Buyel, J. F., With Contri.By, Cajic, S., With Contri.By, Callewaert, N., With Contri.By, Can, Ö., With Contri.By, Cox, M. M. J., With Contri.By, DeLisa, M. P., With Contri.By, De Wachter, C., With Contri.By, Elling, L., With Contri.By, Fischer, R., With Contri.By, Flitsch, S. L., With Contri.By, Frohnmeyer, H., With Contri.By, Gottschalk, J., With Contri.By, Goyette-Desjardins, G., With Contri.By, Heffner, K. M., With Contri.By, Hizal, D. B., With Contri.By, Hoffmann, M., With Contri.By, Holland, T., With Contri.By, Jefferis, R., With Contri.By, Kolarich, D., With Contri.By, Laaf, D., With Contri.By, Lepenies, B., With Contri.By, Malik, A., With Contri.By, Martinez, J. E. R., With Contri.By, Mills, D. C., With Contri.By, Moeller, T. D., With Contri.By, Palomares, L. A., With Contri.By, Pralow, A., With Contri.By, RamÚrez, O. T., With Contri.By, Rapp, E., With Contri.By, Reichl, U., With Contri.By, Rexer, T., With Contri.By, Sack, M., With Contri.By, Schillberg, S., With Contri.By, Schön, K., With Contri.By, Seeberger, P. H., With Contri.By, Srivastava, I. K., With Contri.By, Stoger, E., With Contri.By, Thomas, B., With Contri.By, Twyman, R. M., With Contri.By, Van Landuyt, L., With Contri.By, Varón Silva, D., With Contri.By, Wang, Q., With Contri.By, Weyant, K. B., With Contri.By, and Yates, L. E., With Contri.By

Published
2021
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95. Glycoengineering of Mammalian Expression Systems on a Cellular Level

Author

Heffner, Kelley M., Wang, Qiong, Hizal, Deniz Baycin, Can, Özge, Betenbaugh, Michael J., Scheper, Thomas, Series Editor, Belkin, Shimshon, Editorial Board Member, Bley, Thomas, Editorial Board Member, Bohlmann, Jörg, Editorial Board Member, Gu, Man Bock, Editorial Board Member, Hu, Wei-Shou, Editorial Board Member, Mattiasson, Bo, Editorial Board Member, Olsson, Lisbeth, Editorial Board Member, Seitz, Harald, Editorial Board Member, Silva, Ana Catarina, Editorial Board Member, Ulber, Roland, Editorial Board Member, Zeng, An-Ping, Editorial Board Member, Zhong, Jian-Jiang, Editorial Board Member, Zhou, Weichang, Editorial Board Member, Rapp, Erdmann, editor, Reichl, Udo, editor, Alagesan, K., With Contri.By, Betenbaugh, M. J., With Contri.By, Butler, M., With Contri.By, Buyel, J. F., With Contri.By, Cajic, S., With Contri.By, Callewaert, N., With Contri.By, Can, Ö., With Contri.By, Cox, M. M. J., With Contri.By, DeLisa, M. P., With Contri.By, De Wachter, C., With Contri.By, Elling, L., With Contri.By, Fischer, R., With Contri.By, Flitsch, S. L., With Contri.By, Frohnmeyer, H., With Contri.By, Gottschalk, J., With Contri.By, Goyette-Desjardins, G., With Contri.By, Heffner, K. M., With Contri.By, Hizal, D. B., With Contri.By, Hoffmann, M., With Contri.By, Holland, T., With Contri.By, Jefferis, R., With Contri.By, Kolarich, D., With Contri.By, Laaf, D., With Contri.By, Lepenies, B., With Contri.By, Malik, A., With Contri.By, Martinez, J. E. R., With Contri.By, Mills, D. C., With Contri.By, Moeller, T. D., With Contri.By, Palomares, L. A., With Contri.By, Pralow, A., With Contri.By, RamÚrez, O. T., With Contri.By, Rapp, E., With Contri.By, Reichl, U., With Contri.By, Rexer, T., With Contri.By, Sack, M., With Contri.By, Schillberg, S., With Contri.By, Schön, K., With Contri.By, Seeberger, P. H., With Contri.By, Srivastava, I. K., With Contri.By, Stoger, E., With Contri.By, Thomas, B., With Contri.By, Twyman, R. M., With Contri.By, Van Landuyt, L., With Contri.By, Varón Silva, D., With Contri.By, Wang, Q., With Contri.By, Weyant, K. B., With Contri.By, and Yates, L. E., With Contri.By

Published
2021
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96. Glycoproteomics Technologies in Glycobiotechnology

Author

Alagesan, Kathirvel, Hoffmann, Marcus, Rapp, Erdmann, Kolarich, Daniel, Scheper, Thomas, Series Editor, Belkin, Shimshon, Editorial Board Member, Bley, Thomas, Editorial Board Member, Bohlmann, Jörg, Editorial Board Member, Gu, Man Bock, Editorial Board Member, Hu, Wei-Shou, Editorial Board Member, Mattiasson, Bo, Editorial Board Member, Olsson, Lisbeth, Editorial Board Member, Seitz, Harald, Editorial Board Member, Silva, Ana Catarina, Editorial Board Member, Ulber, Roland, Editorial Board Member, Zeng, An-Ping, Editorial Board Member, Zhong, Jian-Jiang, Editorial Board Member, Zhou, Weichang, Editorial Board Member, Rapp, Erdmann, editor, Reichl, Udo, editor, Alagesan, K., With Contri.By, Betenbaugh, M. J., With Contri.By, Butler, M., With Contri.By, Buyel, J. F., With Contri.By, Cajic, S., With Contri.By, Callewaert, N., With Contri.By, Can, Ö., With Contri.By, Cox, M. M. J., With Contri.By, DeLisa, M. P., With Contri.By, De Wachter, C., With Contri.By, Elling, L., With Contri.By, Fischer, R., With Contri.By, Flitsch, S. L., With Contri.By, Frohnmeyer, H., With Contri.By, Gottschalk, J., With Contri.By, Goyette-Desjardins, G., With Contri.By, Heffner, K. M., With Contri.By, Hizal, D. B., With Contri.By, Hoffmann, M., With Contri.By, Holland, T., With Contri.By, Jefferis, R., With Contri.By, Kolarich, D., With Contri.By, Laaf, D., With Contri.By, Lepenies, B., With Contri.By, Malik, A., With Contri.By, Martinez, J. E. R., With Contri.By, Mills, D. C., With Contri.By, Moeller, T. D., With Contri.By, Palomares, L. A., With Contri.By, Pralow, A., With Contri.By, RamÚrez, O. T., With Contri.By, Rapp, E., With Contri.By, Reichl, U., With Contri.By, Rexer, T., With Contri.By, Sack, M., With Contri.By, Schillberg, S., With Contri.By, Schön, K., With Contri.By, Seeberger, P. H., With Contri.By, Srivastava, I. K., With Contri.By, Stoger, E., With Contri.By, Thomas, B., With Contri.By, Twyman, R. M., With Contri.By, Van Landuyt, L., With Contri.By, Varón Silva, D., With Contri.By, Wang, Q., With Contri.By, Weyant, K. B., With Contri.By, and Yates, L. E., With Contri.By

Published
2021
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97. State-of-the-Art Glycomics Technologies in Glycobiotechnology

Author

Pralow, Alexander, Cajic, Samanta, Alagesan, Kathirvel, Kolarich, Daniel, Rapp, Erdmann, Scheper, Thomas, Series Editor, Belkin, Shimshon, Editorial Board Member, Bley, Thomas, Editorial Board Member, Bohlmann, Jörg, Editorial Board Member, Gu, Man Bock, Editorial Board Member, Hu, Wei-Shou, Editorial Board Member, Mattiasson, Bo, Editorial Board Member, Olsson, Lisbeth, Editorial Board Member, Seitz, Harald, Editorial Board Member, Silva, Ana Catarina, Editorial Board Member, Ulber, Roland, Editorial Board Member, Zeng, An-Ping, Editorial Board Member, Zhong, Jian-Jiang, Editorial Board Member, Zhou, Weichang, Editorial Board Member, Rapp, Erdmann, editor, Reichl, Udo, editor, Alagesan, K., With Contri.By, Betenbaugh, M. J., With Contri.By, Butler, M., With Contri.By, Buyel, J. F., With Contri.By, Cajic, S., With Contri.By, Callewaert, N., With Contri.By, Can, Ö., With Contri.By, Cox, M. M. J., With Contri.By, DeLisa, M. P., With Contri.By, De Wachter, C., With Contri.By, Elling, L., With Contri.By, Fischer, R., With Contri.By, Flitsch, S. L., With Contri.By, Frohnmeyer, H., With Contri.By, Gottschalk, J., With Contri.By, Goyette-Desjardins, G., With Contri.By, Heffner, K. M., With Contri.By, Hizal, D. B., With Contri.By, Hoffmann, M., With Contri.By, Holland, T., With Contri.By, Jefferis, R., With Contri.By, Kolarich, D., With Contri.By, Laaf, D., With Contri.By, Lepenies, B., With Contri.By, Malik, A., With Contri.By, Martinez, J. E. R., With Contri.By, Mills, D. C., With Contri.By, Moeller, T. D., With Contri.By, Palomares, L. A., With Contri.By, Pralow, A., With Contri.By, RamÚrez, O. T., With Contri.By, Rapp, E., With Contri.By, Reichl, U., With Contri.By, Rexer, T., With Contri.By, Sack, M., With Contri.By, Schillberg, S., With Contri.By, Schön, K., With Contri.By, Seeberger, P. H., With Contri.By, Srivastava, I. K., With Contri.By, Stoger, E., With Contri.By, Thomas, B., With Contri.By, Twyman, R. M., With Contri.By, Van Landuyt, L., With Contri.By, Varón Silva, D., With Contri.By, Wang, Q., With Contri.By, Weyant, K. B., With Contri.By, and Yates, L. E., With Contri.By

Published
2021
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98. Impact of Protein Glycosylation on the Design of Viral Vaccines

Author

Schön, Kathleen, Lepenies, Bernd, Goyette-Desjardins, Guillaume, Scheper, Thomas, Series Editor, Belkin, Shimshon, Editorial Board Member, Bley, Thomas, Editorial Board Member, Bohlmann, Jörg, Editorial Board Member, Gu, Man Bock, Editorial Board Member, Hu, Wei-Shou, Editorial Board Member, Mattiasson, Bo, Editorial Board Member, Olsson, Lisbeth, Editorial Board Member, Seitz, Harald, Editorial Board Member, Silva, Ana Catarina, Editorial Board Member, Ulber, Roland, Editorial Board Member, Zeng, An-Ping, Editorial Board Member, Zhong, Jian-Jiang, Editorial Board Member, Zhou, Weichang, Editorial Board Member, Rapp, Erdmann, editor, Reichl, Udo, editor, Alagesan, K., With Contri.By, Betenbaugh, M. J., With Contri.By, Butler, M., With Contri.By, Buyel, J. F., With Contri.By, Cajic, S., With Contri.By, Callewaert, N., With Contri.By, Can, Ö., With Contri.By, Cox, M. M. J., With Contri.By, DeLisa, M. P., With Contri.By, De Wachter, C., With Contri.By, Elling, L., With Contri.By, Fischer, R., With Contri.By, Flitsch, S. L., With Contri.By, Frohnmeyer, H., With Contri.By, Gottschalk, J., With Contri.By, Goyette-Desjardins, G., With Contri.By, Heffner, K. M., With Contri.By, Hizal, D. B., With Contri.By, Hoffmann, M., With Contri.By, Holland, T., With Contri.By, Jefferis, R., With Contri.By, Kolarich, D., With Contri.By, Laaf, D., With Contri.By, Lepenies, B., With Contri.By, Malik, A., With Contri.By, Martinez, J. E. R., With Contri.By, Mills, D. C., With Contri.By, Moeller, T. D., With Contri.By, Palomares, L. A., With Contri.By, Pralow, A., With Contri.By, RamÚrez, O. T., With Contri.By, Rapp, E., With Contri.By, Reichl, U., With Contri.By, Rexer, T., With Contri.By, Sack, M., With Contri.By, Schillberg, S., With Contri.By, Schön, K., With Contri.By, Seeberger, P. H., With Contri.By, Srivastava, I. K., With Contri.By, Stoger, E., With Contri.By, Thomas, B., With Contri.By, Twyman, R. M., With Contri.By, Van Landuyt, L., With Contri.By, Varón Silva, D., With Contri.By, Wang, Q., With Contri.By, Weyant, K. B., With Contri.By, and Yates, L. E., With Contri.By

Published
2021
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99. Glycan Array Technology

Author

Martinez, Juana Elizabeth Reyes, Thomas, Baptiste, Flitsch, Sabine Lahja, Scheper, Thomas, Series Editor, Belkin, Shimshon, Editorial Board Member, Bley, Thomas, Editorial Board Member, Bohlmann, Jörg, Editorial Board Member, Gu, Man Bock, Editorial Board Member, Hu, Wei-Shou, Editorial Board Member, Mattiasson, Bo, Editorial Board Member, Olsson, Lisbeth, Editorial Board Member, Seitz, Harald, Editorial Board Member, Silva, Ana Catarina, Editorial Board Member, Ulber, Roland, Editorial Board Member, Zeng, An-Ping, Editorial Board Member, Zhong, Jian-Jiang, Editorial Board Member, Zhou, Weichang, Editorial Board Member, Rapp, Erdmann, editor, Reichl, Udo, editor, Alagesan, K., With Contri.By, Betenbaugh, M. J., With Contri.By, Butler, M., With Contri.By, Buyel, J. F., With Contri.By, Cajic, S., With Contri.By, Callewaert, N., With Contri.By, Can, Ö., With Contri.By, Cox, M. M. J., With Contri.By, DeLisa, M. P., With Contri.By, De Wachter, C., With Contri.By, Elling, L., With Contri.By, Fischer, R., With Contri.By, Flitsch, S. L., With Contri.By, Frohnmeyer, H., With Contri.By, Gottschalk, J., With Contri.By, Goyette-Desjardins, G., With Contri.By, Heffner, K. M., With Contri.By, Hizal, D. B., With Contri.By, Hoffmann, M., With Contri.By, Holland, T., With Contri.By, Jefferis, R., With Contri.By, Kolarich, D., With Contri.By, Laaf, D., With Contri.By, Lepenies, B., With Contri.By, Malik, A., With Contri.By, Martinez, J. E. R., With Contri.By, Mills, D. C., With Contri.By, Moeller, T. D., With Contri.By, Palomares, L. A., With Contri.By, Pralow, A., With Contri.By, RamÚrez, O. T., With Contri.By, Rapp, E., With Contri.By, Reichl, U., With Contri.By, Rexer, T., With Contri.By, Sack, M., With Contri.By, Schillberg, S., With Contri.By, Schön, K., With Contri.By, Seeberger, P. H., With Contri.By, Srivastava, I. K., With Contri.By, Stoger, E., With Contri.By, Thomas, B., With Contri.By, Twyman, R. M., With Contri.By, Van Landuyt, L., With Contri.By, Varón Silva, D., With Contri.By, Wang, Q., With Contri.By, Weyant, K. B., With Contri.By, and Yates, L. E., With Contri.By

Published
2021
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100. Correction to: Animal Cell Expression Systems

Author

Butler, M., Reichl, U., Scheper, Thomas, Series Editor, Belkin, Shimshon, Editorial Board Member, Bley, Thomas, Editorial Board Member, Bohlmann, Jörg, Editorial Board Member, Gu, Man Bock, Editorial Board Member, Hu, Wei-Shou, Editorial Board Member, Mattiasson, Bo, Editorial Board Member, Olsson, Lisbeth, Editorial Board Member, Seitz, Harald, Editorial Board Member, Silva, Ana Catarina, Editorial Board Member, Ulber, Roland, Editorial Board Member, Zeng, An-Ping, Editorial Board Member, Zhong, Jian-Jiang, Editorial Board Member, Zhou, Weichang, Editorial Board Member, Rapp, Erdmann, editor, Reichl, Udo, editor, Alagesan, K., With Contri.By, Betenbaugh, M. J., With Contri.By, Butler, M., With Contri.By, Buyel, J. F., With Contri.By, Cajic, S., With Contri.By, Callewaert, N., With Contri.By, Can, Ö., With Contri.By, Cox, M. M. J., With Contri.By, DeLisa, M. P., With Contri.By, De Wachter, C., With Contri.By, Elling, L., With Contri.By, Fischer, R., With Contri.By, Flitsch, S. L., With Contri.By, Frohnmeyer, H., With Contri.By, Gottschalk, J., With Contri.By, Goyette-Desjardins, G., With Contri.By, Heffner, K. M., With Contri.By, Hizal, D. B., With Contri.By, Hoffmann, M., With Contri.By, Holland, T., With Contri.By, Jefferis, R., With Contri.By, Kolarich, D., With Contri.By, Laaf, D., With Contri.By, Lepenies, B., With Contri.By, Malik, A., With Contri.By, Martinez, J. E. R., With Contri.By, Mills, D. C., With Contri.By, Moeller, T. D., With Contri.By, Palomares, L. A., With Contri.By, Pralow, A., With Contri.By, RamÚrez, O. T., With Contri.By, Rapp, E., With Contri.By, Reichl, U., With Contri.By, Rexer, T., With Contri.By, Sack, M., With Contri.By, Schillberg, S., With Contri.By, Schön, K., With Contri.By, Seeberger, P. H., With Contri.By, Srivastava, I. K., With Contri.By, Stoger, E., With Contri.By, Thomas, B., With Contri.By, Twyman, R. M., With Contri.By, Van Landuyt, L., With Contri.By, Varón Silva, D., With Contri.By, Wang, Q., With Contri.By, Weyant, K. B., With Contri.By, and Yates, L. E., With Contri.By

Published
2021
Full Text
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