Literature

You will find more information about the specific chapters in following papers.

pdf for download

InVivo´s research results

Poster presentations
  • Beckmann, T.F., Klausing, S., Püngel. S., Coronel, J., Welsink, T., Heinrich, C. 2015. „Medium development beyond production media: Chemically defined media for transfection and single cell cultivation“ BMC Proceedings 9 (Suppl 9) P27 [pdf]
  • Püngel, S., Veiczi, M., Welsink, T., Faust, D., Vater, V., Levison, D., Moeller, U., Weglöhner, W. 2013. „INVect: A Novel Polycationic Reagent For Transient Transfection Of Mammalian Cells“ BMC Proceedings, 7 (Suppl 6):P26 [pdf]
  • Püngel, S., Welsink, T., Villegas Soto, P., Weglöhner, W., Beckmann, T.F., Eickmeier, I., Northoff, S., Heinrich, C. 2013. „Development of a chemically defined cultivation and transfection medium for HEK cell lines“ BMC Proceedings 2013, 7 (Suppl 6):P27 [pdf]
  • Püngel, S., Veiczi, M., Beckmann, T.F., Vater, V., Villegas Soto, P., Ermerling, R., Levison, D., Heinrich, C., Welsink, T. 2015. „Reconciling pillars of transient gene expression: From DNA prep via media, reagent and cell line development to holistic process optimization“ BMC Proceedings, 9 (Suppl 9):P18 [pdf]
Poster
  • Beckmann, T.F., Klausing, S., Püngel. S., Coronel, J., Welsink, T., Northoff, S., Heinrich, C. 2015. „Medium development beyond production media: Chemically defined media for transfection and single cell cutivation“ 24th ESACT Meeting, Barcelona, Spain [Poster]
  • Beckmann, T.F., Klausing, S., Püngel. S., Northoff, S., Büntemeyer, H., Weglöhner, W., Heinrich, C. 2014. „Towards a single chemically defined medium for combining transfection and cultivation of HEK and CHO cell lines“ Cell Culture Engineering XIV, Quebec City, Canada [Poster]
  • Beckmann, T.F., Püngel. S., Klausing, S., Welsink, T., Heinrich, C. 2014. „Towards a single chemically defined medium for combining transfection and cultivation of mammalian cell lines“ 27th Annual Meeting of Japanese Association for Animal Cell Technology, Kitakyushu, Japan [Poster]
  • Püngel, S., Vater, V., Villegas Soto, P., Noll, T., Heinrich, C., Welsink, T. 2015. „Cutting-edge transient gene expression: from dna prep via media, reagent and cell line development to holistic process optimization“ 12th Conference on Protein Expression in Animal Cells, San Diego, California, USA [Poster]
  • Püngel, S., Veiczi, M., Welsink, T., Faust, D., Vater, V., Levison, D., Moeller, U., Weglöhner, W. 2013. „INVect: A Novel Polycationic Reagent For Transient Transfection Of Mammalian Cells“ 23rd ESACT Meeting, Lille, France [Poster]
  • Püngel, S., Welsink, T., Villegas Soto, P., Weglöhner, W., Beckmann, T.F., Eickmeier, I., Northoff, S., Heinrich, C. 2013. „Development of a chemically defined cultivation and transfection medium for HEK cell lines“ 23rd ESACT Meeting, Lille, France [Poster]
  • Levison, D., Moeller, U., Brodersen, N., Welsink, T., Kirchner, S., Goldmann, A., Leister, P., Weglöhner, W. 2011. „Development of a Cationic Reagent for Transient Transfection of Mammalian Cells“ 22nd ESACT Meeting, Vienna, Austria [Poster]
  • Welsink, T., Forke, A., Kirchner, S., Kleinschmidt, A., Goldmann, A., Leister, P., Weglöhner, W. 2011. „Transient Transfection of Human CAP-T Cells – Basics and Optimization“ 22nd ESACT Meeting, Vienna, Austria [Poster]
  • Veiczi, M., Puengel, S., Welsink, T., Faust, D., Vater, V., Weglöhner, W., Levison, D., Moeller, U. 2013. „Development of a Polycationic Reagent for Transient Transfection of Mammalian Cells“ 3rd European Congress of Applied Biotechnology, The Hague, Netherlands [Poster]
Case Studies
  • “Converting rabbit hybridoma into recombinant antibodies with effective transient production in an optimized human expression system”, presented by Dr. T. Welsink at HAH Belfast 2016 [Case studies_HAH Belfast 2016.pdf]
  • Sebastian Püngel, Vanessa Vater, Penélope Villegas Soto and Tim Welsink. 2017. “Expression of Difficult-to-Express (DTE) Proteins” [pdf]
  • Sebastian Püngel, Vanessa Vater, Penélope Villegas Soto and Tim Welsink. 2017. “Expression of recombinant proteins in early and late stage discovery” [pdf]

Literature Overview

Introductory and historical literature
  • Akinc, Akin, Mini Thomas, Alexander M. Klibanov, and Robert Langer. 2005. “Exploring polyethylenimine-mediated DNA transfection and the proton sponge hypothesis.” J Gene Med 7(5):657-663. [PubMed]
  • Backliwal, Gaurav, Markus Hildinger, Vivek Hasija, and Florian M. Wurm. 2008. “High-density transfection with HEK-293 cells allows doubling of transient titers and removes need for a priori DNA complex formation with PEI.” Biotechnol Bioeng 99(3):721-727. [PubMed]
  • Baldi, Lucia, David L. Hacker, Myriam Adam, and Florian M. Wurm. 2007. “Recombinant protein production by large-scale transient gene expression in mammalian cells: state of the art and future perspectives.” Biotechnol Lett 29(5):677-684. [PubMed]
  • Baldi, Lucia, David L. Hacker, Carine Meerschman, and Florian M. Wurm. 2012. “Large-scale transfection of mammalian cells.” Methods Mol Biol 801:13-26. [PubMed]
  • Baldi, Lucia, Natalie Muller, Sylviane Picasso, Raymond Jacquet, Philippe Girard, Huy P. Thanh, Elisabeth Derow, and Florian M. Wurm. 2005. “Transient gene expression in suspension HEK-293 cells: application to large-scale protein production.” Biotechnol Prog 21(1):148-153. [PubMed]
  • Brunner, S., T. Sauer, S. Carotta, M. Cotten, M. Saltik, and E. Wagner. 2000. “Cell cycle dependence of gene transfer by lipoplex, polyplex and recombinant adenovirus.” Gene Ther 7(5):401-407. [PubMed]
  • Cain, Katharine, Shirley Peters, Hanna Hailu, Bernie Sweeney, Paul Stephens, James Heads, Kaushik Sarkar, Andy Ventom, Catherine Page, and Alan Dickson. 2013. “A CHO cell line engineered to express XBP1 and ERO1-Lα has increased levels of transient protein expression.” Biotechnol Prog 29(3):697-706. [PubMed]
  • Carpentier, Eric, Sébastien Paris, Amine A. Kamen, and Yves Durocher. 2007. “Limiting factors governing protein expression following polyethylenimine-mediated gene transfer in HEK293-EBNA1 cells.” J Biotechnol 128(2):268-280. [PubMed]
  • Chahal, Parminder S., Erica Schulze, Rosa Tran, Johnny Montes, and Amine A. Kamen. 2014. “Production of adeno-associated virus (AAV) serotypes by transient transfection of HEK293 cell suspension cultures for gene delivery.” J Virol Methods 196:163-173. [PubMed]
  • Daramola, Olalekan, Jessica Stevenson, Greg Dean, Diane Hatton, Gary Pettman, William Holmes, and Ray Field. 2014. “A high-yielding CHO transient system: coexpression of genes encoding EBNA-1 and GS enhances transient protein expression.” Biotechnol Prog 30(1):132-141. [PubMed]
  • Derouazi, Madiha, Philippe Girard, Frédéric Van Tilborgh, Keyvan Iglesias, Natalie Muller, Martin Bertschinger, and Florian M. Wurm. 2004. “Serum-free large-scale transient transfection of CHO cells.” Biotechnol Bioeng 87(4):537-545. [PubMed]
  • Durocher, Yves, Sylvie Perret, and Amine Kamen. 2002. “High-level and high-throughput recombinant protein production by transient transfection of suspension-growing human 293-EBNA1 cells.” Nucleic Acids Res 30(2):E9. [PubMed]
  • Fliedl, Lukas and Christian Kaisermayer. 2011. “Transient gene expression in HEK293 and vero cells immobilised on microcarriers.” J Biotechnol 153(1-2):15-21. [PubMed]
  • Fliedl, Lukas and Christian Kaisermayer. 2014. “Scalable transient gene expression in adherent mammalian cells using polyethylenimine.” Methods Mol Biol 1104:29-34. [PubMed]
  • Geisse, Sabine. 2009. “Reflections on more than 10 years of TGE approaches.” Protein Expr Purif 64(2):99-107. [PubMed]
  • Geisse, Sabine and Cornelia Fux. 2009. “Recombinant protein production by transient gene transfer into Mammalian cells.” Methods Enzymol 463:223-238. [PubMed]
  • Geisse, Sabine and Mario Henke. 2005. “Large-scale transient transfection of mammalian cells: a newly emerging attractive option for recombinant protein production.” J Struct Funct Genomics 6(2-3):165-170. [PubMed]
  • Geisse, Sabine, Martin Jordan, and Florian M. Wurm. 2005. “Large-scale transient expression of therapeutic proteins in mammalian cells.” Methods Mol Biol 308:87-98. [PubMed]
  • Geisse, Sabine and Bernd Voedisch. 2012. “Transient expression technologies: past, present, and future.” Methods Mol Biol 899:203-219. [PubMed]
  • Girard, Philippe, Madiha Derouazi, Gwendoline Baumgartner, Michaela Bourgeois, Martin Jordan, Barbara Jacko, and Florian M. Wurm. 2002. “100-liter transient transfection.” Cytotechnology 38(1-3):15-21. [PubMed]
  • Hacker, David L., Divor Kiseljak, Yashas Rajendra, Sarah Thurnheer, Lucia Baldi, and Florian M. Wurm. 2013. “Polyethyleneimine-based transient gene expression processes for suspension-adapted HEK-293E and CHO-DG44 cells.” Protein Expr Purif 92(1):67-76. [PubMed]
  • Jordan, Martin and Florian Wurm. 2004. “Transfection of adherent and suspended cells by calcium phosphate.” Methods 33(2):136-143. [PubMed]
  • Knibbs, Randall N., Michael Dame, Melissa R. Allen, Yunhong Ding, William J. Hillegas, James Varani, and Lloyd M. Stoolman. 2003. “Sustained High-Yield Production of Recombinant Proteins in transiently Transfected COS-7 Cells Grown on Trimethylamine-Coated (Hillex) Microcarrier Beads.” Biotechnol Prog 19(1):9-13. [PubMed]
  • Liu, Chaoting, Brian Dalby, Weixing Chen, Jennifer M. Kilzer, and Henry C. Chiou. 2008. “Transient transfection factors for high-level recombinant protein production in suspension cultured mammalian cells.” Mol Biotechnol 39(2):141-153. [PubMed]
  • Meissner, P., H. Pick, A. Kulangara, P. Chatellard, K. Friedrich, and F. M. Wurm. 2001. “Transient gene expression: recombinant protein production with suspension-adapted HEK293-EBNA cells.” Biotechnol Bioeng 75(2):197-203. [PubMed]
  • Nallet, Sophie, Luca Fornelli, Simone Schmitt, Julien Parra, Lucia Baldi, Yury O. Tsybin, and Florian M. Wurm. 2012. “Glycan variability on a recombinant IgG antibody transiently produced in HEK-293E cells.” N Biotechnol 29(4):471-476. [PubMed]
  • Pham, Phuong L., Amine Kamen, and Yves Durocher. 2006. “Large-scale transfection of mammalian cells for the fast production of recombinant protein.” Mol Biotechnol 34(2):225-237. [PubMed]
  • Rajendra, Yashas, Divor Kiseljak, Lucia Baldi, David L. Hacker, and Florian M. Wurm. 2011. “A simple high-yielding process for transient gene expression in CHO cells.” J Biotechnol 153(1-2):22-26. [PubMed]
  • Rajendra, Yashas, Divor Kiseljak, Lucia Baldi, Florian M. Wurm, and David L. Hacker. 2015. “Transcriptional and post-transcriptional limitations of high-yielding, PEI-mediated transient transfection with CHO and HEK-293E cells.” Biotechnol Prog 31(2):541-549. [PubMed]
  • Raymond, Céline, Roseanne Tom, Sylvie Perret, Pascal Moussouami, Denis L’Abbé, Gilles St-Laurent, and Yves Durocher. 2011. “A simplified polyethylenimine-mediated transfection process for large-scale and high-throughput applications.” Methods 55(1):44-51. [PubMed]
  • Rosser, Mary P., Wei Xia, Steven Hartsell, Michael McCaman, Ying Zhu, Soujuan Wang, Susan Harvey, Peter Bringmann, and Ronald R. Cobb. 2005. “Transient transfection of CHO-K1-S using serum-free medium in suspension: a rapid mammalian protein expression system.” Protein Expr Purif 40(2):237-243. [PubMed]
  • Ruponen, Marika, Paavo Honkakoski, Seppo Rönkkö, Jukka Pelkonen, Markku Tammi, and Arto Urtti. 2003. “Extracellular and intracellular barriers in non-viral gene delivery.” J Control Release 93(2):213-217. [PubMed]
  • Schlaeger, Ernst-Jürgen and Klaus Christensen. 1999. “Transient gene expression in mammalian cells grown in serum-free suspension culture.” Cytotechnology 30(1-3):71-83. [PubMed]
  • Sun, Xiangming, Hui C. Hia, Peen E. Goh, and Miranda G. S. Yap. 2008. “High-density transient gene expression in suspension-adapted 293 EBNA1 cells.” Biotechnol Bioeng 99(1):108-116. [PubMed]
  • Thompson, Ben C., Camille R. J. Segarra, Olivia L. Mozley, Olalekan Daramola, Ray Field, Peter R. Levison, and David C. James. 2012. “Cell line specific control of polyethylenimine-mediated transient transfection optimized with “Design of experiments” methodology.” Biotechnol Prog 28(1):179-187. [PubMed]
  • Vink, Tom, Maroeska Oudshoorn-Dickmann, Marcel Roza, Jelte-Jan Reitsma, and Rob N. de Jong. 2013. “A simple, robust and highly efficient transient expression system for producing antibodies.” Methods. [PubMed]
  • Ye, Jianxin, Vanessa Kober, Melanie Tellers, Zubia Naji, Peter Salmon, and Julia F. Markusen. 2009. “High-level protein expression in scalable CHO transient transfection.” Biotechnol Bioeng 103(3):542-551. [PubMed]
  • Zhang, Jianbing, Xin Liu, Andrea Bell, Rebecca To, Toya N. Baral, Ali Azizi, Jianjun Li, Brian Cass, and Yves Durocher. 2009. “Transient expression and purification of chimeric heavy chain antibodies.” Protein Expr Purif 65(1):77-82. [PubMed]
  • Andrew R. M. Bradburya, Nathan D. Trinkleinb, Holger Thie c, Ian C. Wilkinsond, Atul K. Tandone, Stephen Andersond, Catherine L. Bladend, Brittany Jonese, Shelley Force Aldredb, Marco Bestagnof, Oscar Burronef, Jennifer Maynardg, Fortunato Ferraraa, James S. Trimmer h, Janina G€ornemanni, Jacob Glanvillej, Philipp Wolfk, Andre Frenzell,p, Julin Wongm, Xin Yu Kohm, Hui-Yan Engm, David Lanem, Marie-Paule Lefrancn, Mike Clarko, and Stefan Dübel. 2018. „When monoclonal antibodies are not monospecific: Hybridomas frequently express additional functional variable regions“ MABS, Vol. 10, No.4, 539-546 [pdf]
Process Optimization
  • Abbott, Mark W., Brian Middleton, Fredrik Kartberg, Josefine Claesson, Robert Roth, and David Fisher. 2015. “Optimisation of a simple method to transiently transfect a CHO cell line in high-throughput and at large scale.” Protein Expr Purif 116:113-119. [PubMed]
  • Allen, Martin J., James P. Boyce, Michael T. Trentalange, David L. Treiber, Brian Rasmussen, Benjamin Tillotson, Raymond Davis, and Pranhitha Reddy. 2008. “Identification of novel small molecule enhancers of protein production by cultured mammalian cells.” Biotechnol Bioeng 100(6):1193-1204. [PubMed]
  • Backliwal, Gaurav, Markus Hildinger, Ivan Kuettel, Fanny Delegrange, David L. Hacker, and Florian M. Wurm. 2008. “Valproic acid: a viable alternative to sodium butyrate for enhancing protein expression in mammalian cell cultures.” Biotechnol Bioeng 101(1):182-189. [PubMed]
  • Barnard, Gavin C., Maria D. Hougland, and Yashas Rajendra. 2015. “High-throughput mAb expression and purification platform based on transient CHO.” Biotechnol Prog 31(1):239-247. [PubMed]
  • Bollin, Frederic, Vincent Dechavanne, and Laurent Chevalet. 2011. “Design of Experiment in CHO and HEK transient transfection condition optimization.” Protein Expr Purif 78(1):61-68. [PubMed]
  • Bos, Aaron B., Peng Luan, Joseph N. Duque, Dorothea Reilly, Peter D. Harms, and Athena W. Wong. 2015. “Optimization and automation of an end-to-end high throughput microscale transient protein production process.” Biotechnol Bioeng 112(9):1832-1842. [PubMed]
  • Cervera, Laura, Javier Fuenmayor, Irene González-Domínguez, Sonia Gutiérrez-Granados, Maria M. Segura, and Francesc Gòdia. 2015. “Selection and optimization of transfection enhancer additives for increased virus-like particle production in HEK293 suspension cell cultures.” Appl Microbiol Biotechnol 99(23):9935-9949. [PubMed]
  • Cervera, Laura, Sonia Gutiérrez-Granados, Nicholas S. Berrow, Maria M. Segura, and Francesc Gòdia. 2015. “Extended gene expression by medium exchange and repeated transient transfection for recombinant protein production enhancement.” Biotechnol Bioeng 112(5):934-946. [PubMed]
  • de Los Milagros Bassani Molinas, Maria, Christiane Beer, Friedemann Hesse, Manfred Wirth, and Roland Wagner. 2014. “Optimizing the transient transfection process of HEK-293 suspension cells for protein production by nucleotide ratio monitoring.” Cytotechnology 66(3):493-514. [PubMed]
  • Estes, Bram, Yueh-Rong Hsu, Lei-Ting Tam, Jackie Sheng, Jennitte Stevens, and Raj Haldankar. 2015. “Uncovering methods for the prevention of protein aggregation and improvement of product quality in a transient expression system.” Biotechnol Prog 31(1):258-267. [PubMed]
  • Halff, Els F., Marian Versteeg, T. Harma C. Brondijk, and Eric G. Huizinga. 2014. “When less becomes more: optimization of protein expression in HEK293-EBNA1 cells using plasmid titration – a case study for NLRs.” Protein Expr Purif 99:27-34. [PubMed]
  • Hopkins, Ralph F., Vanessa E. Wall, and Dominic Esposito. 2012. “Optimizing transient recombinant protein expression in mammalian cells.” Methods Mol Biol 801:251-268. [PubMed]
  • Jesus, Maria J. D., Philippe Girard, Michaela Bourgeois, Gwendoline Baumgartner, Barbara Jacko, Hanspeter Amstutz, and Florian M. Wurm. 2004. “TubeSpin satellites: a fast track approach for process development with animal cells using shaking technology.” Biochem Eng J 17(3):217-223.
  • Kichler, Antoine, Christian Leborgne, and Olivier Danos. 2005. “Dilution of reporter gene with stuffer DNA does not alter the transfection efficiency of polyethylenimines.” J Gene Med 7(11):1459-1467. [PubMed]
  • Kiseljak, Divor, Yashas Rajendra, Sagar S. Manoli, Lucia Baldi, David L. Hacker, and Florian M. Wurm. 2011. “The use of filler DNA for improved transfection and reduced DNA needs in transient gene expression with CHO and HEK cells.” BMC Proc 5 Suppl 8:P33. [PubMed]
  • Muller, Natalie, Madiha Derouazi, Frédéric Van Tilborgh, Sarah Wulhfard, David L. Hacker, Martin Jordan, and Florian M. Wurm. 2007. “Scalable transient gene expression in Chinese hamster ovary cells in instrumented and non-instrumented cultivation systems.” Biotechnol Lett 29(5):703-711. [PubMed]
  • Parham, J. H., M. A. Iannone, L. K. Overton, and J. T. Hutchins. 1998. “Optimization of transient gene expression in mammalian cells and potential for scale-up using flow electroporation.” Cytotechnology 28(1-3):147-155. [PubMed]
  • Pereira, João, Yashas Rajendra, Lucia Baldi, David L. Hacker, and Florian M. Wurm. 2011. “Transient gene expression with CHO cells in conditioned medium: a study using TubeSpin bioreactors.” BMC Proc 5 Suppl 8:P38. [PubMed]
  • Pham, Phuong L., Sylvie Perret, Brian Cass, Eric Carpentier, Gilles St-Laurent, Louis Bisson, Amine Kamen, and Yves Durocher. 2005. “Transient gene expression in HEK293 cells: peptone addition posttransfection improves recombinant protein synthesis.” Biotechnol Bioeng 90(3):332-344. [PubMed]
  • Pham, Phuong L., Sylvie Perret, Huyen C. Doan, Brian Cass, Gilles St-Laurent, Amine Kamen, and Yves Durocher. 2003. “Large-scale transient transfection of serum-free suspension-growing HEK293 EBNA1 cells: peptone additives improve cell growth and transfection efficiency.” Biotechnol Bioeng 84(3):332-342. [PubMed]
  • Pradhan, Ketaki and Mugdha Gadgil. 2012. “Effect of addition of ‘carrier’ DNA during transient protein expression in suspension CHO culture.” Cytotechnology 64(6):613-22. [PubMed]
  • Rajendra, Yashas, Sowmya Balasubramanian, Divor Kiseljak, Lucia Baldi, Florian M. Wurm, and David L. Hacker. 2015. “Enhanced plasmid DNA utilization in transiently transfected CHO-DG44 cells in the presence of polar solvents.” Biotechnol Prog 31(6):1571-1578. [PubMed]
  • Rajendra, Yashas, Divor Kiseljak, Lucia Baldi, David L. Hacker, and Florian M. Wurm. 2012. “Reduced glutamine concentration improves protein production in growth-arrested CHO-DG44 and HEK-293E cells.” Biotechnol Lett 34(4):619-626. [PubMed]
  • Rajendra, Yashas, Divor Kiseljak, Sagar Manoli, Lucia Baldi, David L. Hacker, and Florian M. Wurm. 2012. “Role of non-specific DNA in reducing coding DNA requirement for transient gene expression with CHO and HEK-293E cells.” Biotechnol Bioeng 109(9):2271-2278. [PubMed]
  • Sun, Xiangming, Peen E. Goh, Kathy T. K. Wong, Tetsushi Mori, and Miranda G. S. Yap. 2006. “Enhancement of transient gene expression by fed-batch culture of HEK 293 EBNA1 cells in suspension.” Biotechnol Lett 28(11):843-848. [PubMed]
  • Thompson, Ben C., Camille R. J. Segarra, Olivia L. Mozley, Olalekan Daramola, Ray Field, Peter R. Levison, and David C. James. 2012. “Cell line specific control of polyethylenimine-mediated transient transfection optimized with “Design of experiments” methodology.” Biotechnol Prog 28(1):179-187. [PubMed]
  • Tuvesson, Ola, Christina Uhe, Aleksei Rozkov, and Elke Lüllau. 2008. “Development of a generic transient transfection process at 100 L scale.” Cytotechnology 56(2):123-136. [PubMed]
  • Vink, Tom, Maroeska Oudshoorn-Dickmann, Marcel Roza, Jelte-Jan Reitsma, and Rob N. de Jong. 2014. “A simple, robust and highly efficient transient expression system for producing antibodies.” Methods 65(1):5-10. [PubMed]
  • Wulhfard, Sarah, Lucia Baldi, David L. Hacker, and Florian Wurm. 2010. “Valproic acid enhances recombinant mRNA and protein levels in transiently transfected Chinese hamster ovary cells.” J Biotechnol 148(2-3):128-132. [PubMed]
  • Wulhfard, Sarah, Stéphanie Tissot, Sophie Bouchet, Jean Cevey, Maria D. Jesus, David L. Hacker, and Florian M. Wurm. 2008. “Mild hypothermia improves transient gene expression yields several fold in Chinese hamster ovary cells.” Biotechnol Prog 24(2):458-465. [PubMed]
  • You, Min, Yanning Liu, Yingwei Chen, Jiyuan Guo, Jun Wu, Yajuan Fu, Rong Shen, Rui Qi, Wenxin Luo, and Ningshao Xia. 2013. “Maximizing antibody production in suspension-cultured mammalian cells by the customized transient gene expression method.” Biosci Biotechnol Biochem 77(6):1207-1213. [PubMed]
  • Zhang, Xiaowei, Isabel F. Garcia, Lucia Baldi, David L. Hacker, and Florian M. Wurm. 2010. “Hyperosmolarity enhances transient recombinant protein yield in Chinese hamster ovary cells.” Biotechnol Lett 32(11):1587-1592. [PubMed]
Cell Culture Media
  • Jan, D. C., S. J. Jones, A. N. Emery, and M. al Rubeai. 1994. “Peptone, a low-cost growth-promoting nutrient for intensive animal cell culture.” Cytotechnology 16(1):17-26. [PubMed]
  • Keenan, J. and M. Clynes. 1996. “Replacement of transferrin by simple iron compounds for MDCK cells grown and subcultured in serum-free medium.” In Vitro Cell Dev Biol Anim 32(8):451-453. [PubMed]
  • Merten, O. W., J. V. Kierulff, N. Castignolles, and P. Perrin. 1994. “Evaluation of the new serum-free medium (MDSS2) for the production of different biologicals: use of various cell lines.” Cytotechnology 14(1):47-59. [PubMed]
  • Mizrahi, A. and A. Lazar. 1988. “Media for cultivation of animal cells: an overview.” Cytotechnology 1(3):199-214. [PubMed]
  • Qi, Y. M., P. F. Greenfield, and S. Reid. 1996. “Evaluation of a simple protein free medium that supports high levels of monoclonal antibody production.” Cytotechnology 21(2):95-109. [PubMed]
Cell Line
  • Cain, Katharine, Shirley Peters, Hanna Hailu, Bernie Sweeney, Paul Stephens, James Heads, Kaushik Sarkar, Andy Ventom, Catherine Page, and Alan Dickson. 2013. “A CHO cell line engineered to express XBP1 and ERO1-Lα has increased levels of transient protein expression.” Biotechnol Prog 29(3):697-706. [PubMed]
  • Chiou, Henry C., Sanjay Vasu, Chao Y. Liu, Isabel Cisneros, Meredith B. Jones, and Jonathan F. Zmuda. 2014. “Scalable transient protein expression.” Methods Mol Biol 1104:35-55. [PubMed]
  • Codamo, Joe, Trent P. Munro, Benjamin S. Hughes, Michael Song, and Peter P. Gray. 2011. “Enhanced CHO cell-based transient gene expression with the epi-CHO expression system.” Mol Biotechnol 48(2):109-115. [PubMed]
  • Fischer, Simon, Nadine Charara, Andrea Gerber, Jens Wölfel, Gudrun Schiedner, Bernd Voedisch, and Sabine Geisse. 2012. “Transient recombinant protein expression in a human amniocyte cell line: the CAP-T® cell system.” Biotechnol Bioeng 109(9):2250-2261. [PubMed]
  • Hacker, David L., Divor Kiseljak, Yashas Rajendra, Sarah Thurnheer, Lucia Baldi, and Florian M. Wurm. 2013. “Polyethyleneimine-based transient gene expression processes for suspension-adapted HEK-293E and CHO-DG44 cells.” Protein Expr Purif 92(1):67-76. [PubMed]
  • Kunaparaju, Rajkumar, Mimi Liao, and Noelle-Anne Sunstrom. 2005. “Epi-CHO, an episomal expression system for recombinant protein production in CHO cells.” Biotechnol Bioeng 91(6):670-677. [PubMed]
  • Liu, Chaoting, Brian Dalby, Weixing Chen, Jennifer M. Kilzer, and Henry C. Chiou. 2008. “Transient transfection factors for high-level recombinant protein production in suspension cultured mammalian cells.” Mol Biotechnol 39(2):141-153. [PubMed]
  • Macaraeg, Nichole F., Dorothea E. Reilly, and Athena W. Wong. 2013. “Use of an anti-apoptotic CHO cell line for transient gene expression.” Biotechnol Prog 29(4):1050-1058. [PubMed]
  • Majors, Brian S., Michael J. Betenbaugh, Nels E. Pederson, and Gisela G. Chiang. 2008. “Enhancement of transient gene expression and culture viability using Chinese hamster ovary cells overexpressing Bcl-x(L).” Biotechnol Bioeng 101(3):567-578. [PubMed]
  • Majors, Brian S., Gisela G. Chiang, and Michael J. Betenbaugh. 2009. “Protein and genome evolution in Mammalian cells for biotechnology applications.” Mol Biotechnol 42(2):216-223. [PubMed]
  • Rajendra, Yashas, Maria D. Hougland, Riazul Alam, Teresa A. Morehead, and Gavin C. Barnard. 2015. “A high cell density transient transfection system for therapeutic protein expression based on a CHO GS-knockout cell line: process development and product quality assessment.” Biotechnol Bioeng 112(5):977-986. [PubMed]
  • Strotbek, Michaela, Lore Florin, Jennifer Koenitzer, Anne Tolstrup, Hitto Kaufmann, Angelika Hausser, and Monilola A. Olayioye. 2013. “Stable microRNA expression enhances therapeutic antibody productivity of Chinese hamster ovary cells.” Metab Eng 20:157-166. [PubMed]
  • Zustiak, Matthew P., Lisa Jose, Yueqing Xie, Jianwei Zhu, and Michael J. Betenbaugh. 2014. “Enhanced transient recombinant protein production in CHO cells through the co-transfection of the product gene with Bcl-xL.” Biotechnol J 9(9):1164-1174. [PubMed]
Transfection Reagent
  • Akinc, Akin, Mini Thomas, Alexander M. Klibanov, and Robert Langer. 2005. “Exploring polyethylenimine-mediated DNA transfection and the proton sponge hypothesis.” J Gene Med 7(5):657-663. [PubMed]
  • Asayama, Shoichiro, Ai Hamaya, Takashi Sekine, Hiroyoshi Kawakami, and Shohji Nagaoka. 2004. “Aminated poly(L-histidine) as new pH-sensitive DNA carrier.” Nucleic Acids Symp Ser (Oxf) (48):229-230. [PubMed]
  • Bertschinger, Martin, Gaurav Backliwal, Arnaud Schertenleib, Martin Jordan, David L. Hacker, and Florian M. Wurm. 2006. “Disassembly of polyethylenimine-DNA particles in vitro: implications for polyethylenimine-mediated DNA delivery.” J Control Release 116(1):96-104. [PubMed]
  • Bertschinger, Martin, Sophie Chaboche, Martin Jordan, and Florian M. Wurm. 2004. “A spectrophotometric assay for the quantification of polyethylenimine in DNA nanoparticles.” Anal Biochem 334(1):196-198. [PubMed]
  • Bertschinger, Martin, Arnaud Schertenleib, Jean Cevey, David L. Hacker, and Florian M. Wurm. 2008. “The kinetics of polyethylenimine-mediated transfection in suspension cultures of Chinese hamster ovary cells.” Mol Biotechnol 40(2):136-143. [PubMed]
  • Boussif, O., F. Lezoualc’h, M.A. Zanta, M.D. Mergny, D. Scherman, B. Demeneix, and J.P. Behr. 1995. “A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine.” Proc Natl Acad Sci U S A 92(16):7297–7301. [PubMed]
  • Breunig, Miriam, Uta Lungwitz, Renate Liebl, Claudia Fontanari, Juergen Klar, Armin Kurtz, Torsten Blunk, and Achim Goepferich. 2005. “Gene delivery with low molecular weight linear polyethylenimines.” J Gene Med 7(10):1287-1298. [PubMed]
  • Breunig, Miriam, Uta Lungwitz, Renate Liebl, and Achim Goepferich. 2007. “Breaking up the correlation between efficacy and toxicity for nonviral gene delivery.” Proc Natl Acad Sci U S A 104(36):14454-14459. [PubMed]
  • Breunig, Miriam, Uta Lungwitz, Renate Liebl, Juergen Klar, Birgit Obermayer, Torsten Blunk, and Achim Goepferich. 2007. “Mechanistic insights into linear polyethylenimine-mediated gene transfer.” Biochim Biophys Acta 1770(2):196-205. [PubMed]
  • Choosakoonkriang, Sirirat, Brian A. Lobo, Gary S. Koe, Janet G. Koe, and C. R. Middaugh. 2003. “Biophysical characterization of PEI/DNA complexes.” J Pharm Sci 92(8):1710-1722. [PubMed]
  • Fischer, Dagmar, Anke von Harpe, Klaus Kunath, Holger Petersen, Youxin Li, and Thomas Kissel. 2002. “Copolymers of ethylene imine and N-(2-hydroxyethyl)-ethylene imine as tools to study effects of polymer structure on physicochemical and biological properties of DNA complexes.” Bioconjug Chem 13(5):1124-1133. [PubMed]
  • Fukumoto, Yasunori, Yuuki Obata, Kenichi Ishibashi, Naoki Tamura, Ikue Kikuchi, Kazumasa Aoyama, Yasuyuki Hattori, Kunihiko Tsuda, Yuji Nakayama, and Naoto Yamaguchi. 2010. “Cost-effective gene transfection by DNA compaction at pH 4.0 using acidified, long shelf-life polyethylenimine.” Cytotechnology 62(1):73-82. [PubMed]
  • Gillard, Marianne, Zhongfan Jia, Jeff J. C. Hou, Michael Song, Peter P. Gray, Trent P. Munro, and Michael J. Monteiro. 2014. “Intracellular trafficking pathways for nuclear delivery of plasmid DNA complexed with highly efficient endosome escape polymers.” Biomacromolecules 15(10):3569-3576. [PubMed]
  • Godbey, W. T., K. K. Wu, G. J. Hirasaki, and A. G. Mikos. 1999. “Improved packing of poly(ethylenimine)/DNA complexes increases transfection efficiency.” Gene Ther 6(8):1380-1388. [PubMed]
  • Godbey, W. T., K. K. Wu, and A. G. Mikos. 1999. “Tracking the intracellular path of poly(ethylenimine)/DNA complexes for gene delivery.” Proc Natl Acad Sci U S A 96(9):5177-5181. [PubMed]
  • Gosselin, M. A., W. Guo, and R. J. Lee. 2001. “Efficient gene transfer using reversibly cross-linked low molecular weight polyethylenimine.” Bioconjug Chem 12(6):989-994. [PubMed]
  • Han, Xiangzong, Qiangyi Fang, Feng Yao, Xiaoning Wang, Jufang Wang, Shengli Yang, and Bing Q. Shen. 2009. “The heterogeneous nature of polyethylenimine-DNA complex formation affects transient gene expression.” Cytotechnology 60:63-75. [PubMed]
  • Hanzlikova, Martina, Marika Ruponen, Emilia Galli, Atso Raasmaja, Vladimir Aseyev, Heikki Tenhu, Arto Urtti, and Marjo Yliperttula. 2011. “Mechanisms of polyethylenimine-mediated DNA delivery: free carrier helps to overcome the barrier of cell-surface glycosaminoglycans.” J Gene Med 13(7-8):402-409. [PubMed]
  • Kadlecova, Zuzana, Sophie Nallet, David L. Hacker, Lucia Baldi, Harm-Anton Klok, and Florian M. Wurm. 2012. “Poly(ethyleneimine)-mediated large-scale transient gene expression: influence of molecular weight, polydispersity and N-propionyl groups.” Macromol Biosci 12(5):628-636. [PubMed]
  • Kadlecova, Zuzana, Yashas Rajendra, Mattia Matasci, Lucia Baldi, David L. Hacker, Florian M. Wurm, and Harm-Anton Klok. 2013. “DNA delivery with hyperbranched polylysine: A comparative study with linear and dendritic polylysine.” J Control Release 169(3):276-288. [PubMed]
  • Kadlecova, Zuzana, Yashas Rajendra, Mattia Matasci, David Hacker, Lucia Baldi, Florian M. Wurm, and Harm-Anton Klok. 2012. “Hyperbranched polylysine: a versatile, biodegradable transfection agent for the production of recombinant proteins by transient gene expression and the transfection of primary cells.” Macromol Biosci 12(6):794-804. [PubMed]
  • Kichler, Antoine. 2004. “Gene transfer with modified polyethylenimines.” J Gene Med 6 Suppl 1:S3–10. [PubMed]
  • Kunath, Klaus, Anke von Harpe, Dagmar Fischer, Holger Petersen, Ulrich Bickel, Karlheinz Voigt, and Thomas Kissel. 2003. “Low-molecular-weight polyethylenimine as a non-viral vector for DNA delivery: comparison of physicochemical properties, transfection efficiency and in vivo distribution with high-molecular-weight polyethylenimine.” J Control Release 89(1):113-125. [PubMed]
  • Ladewig, Katharina, Zhi P. Xu, Peter Gray, and G. Q. Max Lu. 2014. “Polyethyleneimine-poly(ethylene glycol)-star-copolymers as efficient and biodegradable vectors for mammalian cell transfection.” J Biomed Mater Res A 102(7):2137-2146. [PubMed]
  • Li, Da, Qing qing Wang, Gu ping Tang, Hong liang Huang, Fen ping Shen, Jing zhong Li, and Hai Yu. 2006. “Receptor-mediated gene delivery using polyethylenimine (PEI) coupled with polypeptides targeting FGF receptors on cells surface.” J Zhejiang Univ Sci B 7(11):906-911. [PubMed]
  • Lungwitz, U., M. Breunig, T. Blunk, and A. Göpferich. 2005. “Polyethylenimine-based non-viral gene delivery systems.” Eur J Pharm Biopharm 60(2):247-266. [PubMed]
  • Neu, Michael, Dagmar Fischer, and Thomas Kissel. 2005. “Recent advances in rational gene transfer vector design based on poly(ethylene imine) and its derivatives.” J Gene Med 7(8):992-1009. [PubMed]
  • Pack, Daniel W., Allan S. Hoffman, Suzie Pun, and Patrick S. Stayton. 2005. “Design and development of polymers for gene delivery.” Nat Rev Drug Discov 4(7):581-593. [PubMed]
  • Park, Kinam. 2009. “PEI-DNA complexes with higher transfection efficiency and lower cytotoxicity.” J Control Release 140(1):1. [PubMed]
  • Vasir, Jaspreet K. and Vinod Labhasetwar. 2008. “Preparation of biodegradable nanoparticles and their use in transfection.” CSH Protoc 2008:pdb.prot4888. [PubMed]
  • Wen, Yuting, Shirong Pan, Xin Luo, Xuan Zhang, Wei Zhang, and Min Feng. 2009. “A biodegradable low molecular weight polyethylenimine derivative as low toxicity and efficient gene vector.” Bioconjug Chem 20(2):322-332. [PubMed]
  • Xiang, Shengnan, Haijun Tong, Qin Shi, Julio C. Fernandes, Tuo Jin, Kerong Dai, and Xiaoling Zhang. 2012. “Uptake mechanisms of non-viral gene delivery.” J Control Release 158(3):371-378. [PubMed]
  • Xie, Qiuling, Guo Xinyong, Chen Xianjin, and Wang Yayu. 2013. “PEI/DNA formation affects transient gene expression in suspension Chinese hamster ovary cells via a one-step transfection process.” Cytotechnology 65(2):263-271. [PubMed]
Vector Design
  • Backliwal, Gaurav, Markus Hildinger, Sebastien Chenuet, Maria Dejesus, and Florian M. Wurm. 2008. “Coexpression of acidic fibroblast growth factor enhances specific productivity and antibody titers in transiently transfected HEK293 cells.” N Biotechnol 25(2-3):162-166. [PubMed]
  • Backliwal, Gaurav, Markus Hildinger, Sebastien Chenuet, Sarah Wulhfard, Maria D. Jesus, and Florian M. Wurm. 2008. “Rational vector design and multi-pathway modulation of HEK 293E cells yield recombinant antibody titers exceeding 1 g/l by transient transfection under serum-free conditions.” Nucleic Acids Res 36(15):e96. [PubMed]
  • Codamo, Joe, Trent P. Munro, Benjamin S. Hughes, Michael Song, and Peter P. Gray. 2011. “Enhanced CHO cell-based transient gene expression with the epi-CHO expression system.” Mol Biotechnol 48(2):109-115. [PubMed]
  • Hackl, Matthias and Nicole Borth. 2014. “One plus one makes three: adding value by co-transfection of anti-apoptotic genes during transient gene expression.” Biotechnol J 9(9):1101-1102. [PubMed]
  • Ho, Steven C. L. and Yuansheng Yang. 2014. “Identifying and engineering promoters for high level and sustainable therapeutic recombinant protein production in cultured mammalian cells.” Biotechnol Lett 36(8):1569-1579. [PubMed]
  • Kober, Lars, Christoph Zehe, and Juergen Bode. 2013. “Optimized signal peptides for the development of high expressing CHO cell lines.” Biotechnol Bioeng 110(4):1164-1173. [PubMed]
  • Kunaparaju, Rajkumar, Mimi Liao, and Noelle-Anne Sunstrom. 2005. “Epi-CHO, an episomal expression system for recombinant protein production in CHO cells.” Biotechnol Bioeng 91(6):670-677. [PubMed]
  • Mahon, Matthew J. 2011. “Vectors bicistronically linking a gene of interest to the SV40 large T antigen in combination with the SV40 origin of replication enhance transient protein expression and luciferase reporter activity.” Biotechniques 51(2):119-128. [PubMed]
  • Tigges, Marcel and Martin Fussenegger. 2006. “Xbp1-based engineering of secretory capacity enhances the productivity of Chinese hamster ovary cells.” Metab Eng 8(3):264-272. [PubMed]
  • Xia, Wei, Peter Bringmann, and John McClary. 2006. “High levels of protein expression using different mammalian CMV promoters in several cell lines.” Protein Expr Purif 45(1):115-124. [PubMed]
Plasmid Preparation
  • Cai, Ying, Stephen Rodriguez, Ramu Rameswaran, Ruxandra Draghia-Akli, Robert J. Juba, and Henry Hebel. 2010. “Production of pharmaceutical-grade plasmids at high concentration and high supercoiled percentage.” Vaccine 28(8):2046-2052. [PubMed]
  • Cheng, Lu, Xiangming Sun, Xiaoping Yi, and Yuanxing Zhang. 2011. “Large-scale plasmid preparation for transient gene expression.” Biotechnol Lett 33(8):1559-1564. [PubMed]
  • Ma, Rui, Jun Zhao, Hui-Cong Du, Shuang Tian, and Li-Wen Li. 2012. “Removing endotoxin from plasmid samples by Triton X-114 isothermal extraction.” Anal Biochem 424(2):124-126. [PubMed]
  • Rozkov, Aleksei, Bert Larsson, Stefan Gillström, Robert Björnestedt, and Stefan R. Schmidt. 2008. “Large-scale production of endotoxin-free plasmids for transient expression in mammalian cell culture.” Biotechnol Bioeng 99(3):557-566. [PubMed]
  • Schmid, G., E. J. Schlaeger, and B. Wipf. 2001. “Non-GMP plasmid production for transient transfection in bioreactors.” Cytotechnology 35(3):157-164. [PubMed]
  • Wright, Jason L., Martin Jordan, and Florian M. Wurm. 2001. “Extraction of plasmid DNA using reactor scale alkaline lysis and selective precipitation for scalable transient transfection.” Cytotechnology 35(3):165-173. [PubMed]
  • Wright, J. L., M. Jordan, and F. M. Wurm. 2003. “Transfection of partially purified plasmid DNA for high level transient protein expression in HEK293-EBNA cells.” J Biotechnol 102(3):211-221. [PubMed]

This list is not intended to be exhaustive. Please feel free to contact us if you notice that a publication is missing from this list.

Stay in the loop with our newsletter!

.