Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 429383
Title 3D Domain Swapping Causes Extensive Multimerisation of Human Interleukin-10 When Expressed In Planta
Author(s) Westerhof, L.B.; Wilbers, R.H.P.; Roosien, J.; Velde, J. van de; Goverse, A.; Bakker, J.; Schots, A.
Source PLoS One 7 (2012)10. - ISSN 1932-6203 - 10 p.
Department(s) Laboratory of Nematology
Publication type Refereed Article in a scientific journal
Publication year 2012
Keyword(s) crystal-structure - transgenic tobacco - escherichia-coli - interferon-gamma - receptor - reveals - il-10 - csf - disease - cytosol
Abstract Heterologous expression platforms of biopharmaceutical proteins have been significantly improved over the last decade. Further improvement can be established by examining the intrinsic properties of proteins. Interleukin-10 (IL-10) is an anti-inflammatory cytokine with a short half-life that plays an important role in re-establishing immune homeostasis. This homodimeric protein of 36 kDa has significant therapeutic potential to treat inflammatory and autoimmune diseases. In this study we show that the major production bottleneck of human IL-10 is not protein instability as previously suggested, but extensive multimerisation due to its intrinsic 3D domain swapping characteristic. Extensive multimerisation of human IL-10 could be visualised as granules in planta. On the other hand, mouse IL-10 hardly multimerised, which could be largely attributed to its glycosylation. By introducing a short glycine-serine-linker between the fourth and fifth alpha helix of human IL-10 a stable monomeric form of IL-10 (hIL-10mono) was created that no longer multimerised and increased yield up to 20-fold. However, hIL-10mono no longer had the ability to reduce pro-inflammatory cytokine secretion from lipopolysaccharide-stimulated macrophages. Forcing dimerisation restored biological activity. This was achieved by fusing human IL-10mono to the C-terminal end of constant domains 2 and 3 of human immunoglobulin A (Fca), a natural dimer. Stable dimeric forms of IL-10, like Fca-IL-10, may not only be a better format for improved production, but also a more suitable format for medical applications.
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