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Gewählte Publikation:

Floss, DM; Sack, M; Arcalis, E; Stadlmann, J; Quendler, H; Rademacher, T; Stoger, E; Scheller, J; Fischer, R; Conrad, U.
(2009): Influence of elastin-like peptide fusions on the quantity and quality of a tobacco-derived human immunodeficiency virus-neutralizing antibody
PLANT BIOTECHNOL J. 2009; 7(9): 899-913. FullText FullText_BOKU

P>The use of vaginal microbicides containing human immunodeficiency virus (HIV)-neutralizing antibodies (nAbs) is a promising strategy to prevent HIV-1 infection. Although antibodies are predominantly manufactured using mammalian cells, elastin-like peptide (ELP) fusion technology improves the stability of recombinant, plant-produced proteins and facilitates their purification, making plants an alternative platform for antibody production. We generated transgenic tobacco plants accumulating four different formats of the anti-HIV-1 antibody 2G12 in the endoplasmic reticulum (ER), i.e. with ELP on either the light or heavy chain, on both, or on neither. Detailed analysis of affinity-purified antibodies by surface plasmon resonance spectroscopy showed that the kinetic binding parameters of all formats were identical to 2G12 lacking ELP produced in Chinese hamster ovary (CHO) cells. Importantly, protein purification from seeds by inverse transition cycling (ITC) did not affect the binding kinetics. Analysis of heavy chain N-glycans from leaf-derived antibodies showed that retrieval to the ER was efficient for all formats. In seeds, however, N-glycans on the naked antibody were extensively trimmed compared with those on the ELP fusion formats, and were localized to a different subcellular compartment. The in vitro HIV-neutralization properties of the tobacco-derived 2G12 were equivalent to or better than those of the CHO counterpart.
Autor*innen der BOKU Wien:
Arcalis Elsa
Quendler Heribert
Stadlmann Johannes
Stöger Eva
BOKU Gendermonitor:

Find related publications in this database (using NML MeSH Indexing)
Animals -
Antibodies, Monoclonal - biosynthesis
Antibodies, Neutralizing - biosynthesis
CHO Cells -
Cricetinae -
Cricetulus -
Elastin - genetics
Endoplasmic Reticulum - metabolism
Gene Expression Regulation, Plant -
Genetic Vectors -
Glycosylation -
HIV Antibodies - biosynthesis
Neutralization Tests -
Plant Leaves - genetics
Plants, Genetically Modified - genetics
Recombinant Fusion Proteins - adverse effects
Seeds - genetics
Tobacco - genetics

Find related publications in this database (Keywords)
human immunodeficiency virus
elastin-like peptide fusion
protein trafficking

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