PMID: 11320504Apr 26, 2001Paper

Characterization of monoclonal antibody fragments produced by plant cells

Biotechnology and Bioengineering
J M Sharp, Pauline M Doran

Abstract

Production of a murine IgG1 was investigated using hairy roots, shooty teratomas, and suspended cells of transgenic tobacco. In all cases, in addition to complete assembled antibody, two to four major antibody fragments accumulated in the biomass. A range of protease inhibitors, protein-stabilizing agents, inhibitors of N-glycosylation and protein secretion, glycan-reactive agents, and affinity probes was used to characterize these fragments and investigate their sites and mechanisms of formation. The fragments were not experimental artifacts caused by antibody degradation during tissue homogenization and sample preparation, nor did they represent glycosylation variants. All of the molecules were actively secreted into the culture media and some showed evidence of Golgi-associated glycan processing, indicating they were not assembly intermediates. Antibody fragments of 50 and 80 kDa were identified mainly as the products of extracellular degradation in the root and shoot apoplast; the 80-kDa fragment was also present in cell suspension medium, and in suspended cell biomass toward the end of the growth phase. Larger 120- and 135-kDa fragments were most likely produced by proteolytic degradation along the secretory pathway outsid...Continue Reading

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Citations

Jan 28, 2003·Vaccine·Fabricio Medina-BolivarCarole L Cramer
Oct 11, 2003·Biotechnology Advances·S R Rao, G A Ravishankar
Apr 13, 2002·Current Opinion in Biotechnology·Dana C Andersen, Lynne Krummen
Apr 16, 2002·Current Opinion in Biotechnology·Eva StogerPaul Christou
Aug 10, 2006·Plant Physiology·Slavko KomarnytskyIlya Raskin
May 10, 2005·Immunology and Cell Biology·M Manuela Rigano, Amanda M Walmsley
Jan 3, 2012·BMC Biotechnology·Verena K HehleCraig J van Dolleweerd
Jun 21, 2011·International Journal of Molecular Sciences·David R ThomasAmanda M Walmsley
Jun 12, 2003·Proceedings of the National Academy of Sciences of the United States of America·Kisung KoHilary Koprowski
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Jul 18, 2015·Bioengineered·Marcello DoniniPatrizio Di Micco
Aug 19, 2011·Biotechnology Advances·Ting-Kuo Huang, Karen A McDonald
Jan 18, 2011·Biotechnology Advances·Jianfeng XuMaureen C Dolan
May 26, 2009·Biotechnology Advances·Sharon M-H Shih, Pauline M Doran
Mar 26, 2016·Frontiers in Plant Science·Manoj K MandalAndreas Schiermeyer
Apr 10, 2007·Protein Expression and Purification·Shin-Young HongMoon-Sik Yang
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May 3, 2008·Plant Biotechnology Journal·Meriem BenchabaneDominique Michaud
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Nov 3, 2006·Protein Expression and Purification·Song-Jae LeeDong-Il Kim
Jul 18, 2006·Trends in Biotechnology·Pauline M Doran
Feb 2, 2006·Journal of Biotechnology·Willemien H WielandDiego V Orzáez

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