Sökning: WFRF:(Dahlén Eva)
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Development of inte...
Development of interleukin-1 receptor antagonist mutants with enhanced antagonistic activity in vitro and improved therapeutic efficacy in collagen-induced arthritis.
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- Dahlén, Eva (författare)
- Lund University,Lunds universitet,Medicinska fakulteten,Faculty of Medicine
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Barchan, Karin (författare)
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Herrlander, Daniel (författare)
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Höjman, Patrik (författare)
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- Karlsson, Marie (författare)
- Lund University,Lunds universitet,Institutionen för experimentell medicinsk vetenskap,Medicinska fakulteten,Department of Experimental Medical Science,Faculty of Medicine
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Ljung, Lill (författare)
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Andersson, Mats (författare)
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Bäckman, Eva (författare)
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- Malmborg Hager, Ann-Christin (författare)
- Lund University,Lunds universitet,Institutionen för immunteknologi,Institutioner vid LTH,Lunds Tekniska Högskola,Department of Immunotechnology,Departments at LTH,Faculty of Engineering, LTH
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Walse, Björn (författare)
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Joosten, Leo (författare)
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van den Berg, Wim (författare)
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(creator_code:org_t)
- 2008-10-09
- 2008
- Engelska.
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Ingår i: Journal of Immunotoxicology. - : Informa UK Limited. - 1547-6901 .- 1547-691X. ; 5:2, s. 189-199
- Relaterad länk:
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http://www.ncbi.nlm....
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http://dx.doi.org/10...
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https://lup.lub.lu.s...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- Interleukin-1 receptor antagonist (IL-1Ra) is a naturally occurring inhibitor of the pro-inflammatory interleukin-1-mediated activation of the interleukin-1 receptor (IL-1R). Although wild-type IL-1Ra is used for treatment of inflammatory diseases, its effect is moderate and/or short-lived. The objective of this study was to generate IL-1Ra mutants with enhanced antagonistic activity for potential therapeutic use. Using a directed evolution approach in which libraries of IL-1Ra gene mutants were generated and screened in functional assays, mutants with desired properties were identified. Initially, diversity was introduced into the IL-1Ra using random mutagenesis. Mutations resulting in enhanced antagonistic activity were identified by screening in a reporter cell assay. To further enhance the antagonistic activity, selected mutations were recombined using the DNA recombination technology Fragment-INduced Diversity (FIND). Following three rounds of FIND recombination, several mutants with up to nine times enhanced antagonistic activity (mean IC50 +/- SEM value: 0.78 +/- 0.050 vs. 6.8 +/- 1.1 ng/ml for mutant and wild-type, respectively) were identified. Sequence analysis identified the mutations D47N, E52R and E90Y as being most important for this effect, however, the mutations P38Y, H54R, Q129L and M136N further enhanced the antagonistic function. Analysis of identified mutations in protein models based on the crystal structure of the IL-1Ra/IL-1R complex suggested that mutations found to enhance the antagonistic activity had a stabilizing effect on the IL-1Ra mutants or increased the affinity for the IL-1R. Finally, the therapeutic effect of one mutant was compared to that of wild-type IL-1Ra in collagen-induced arthritis in mice. Indeed, the enhanced antagonistic effect of the mutants observed in vitro was also seen in vivo. In conclusion, these results demonstrate that directed evolution of IL-1Ra is an effective means of generating highly potent therapeutic proteins.
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- art (ämneskategori)
- ref (ämneskategori)
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Till lärosätets databas
- Av författaren/redakt...
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Dahlén, Eva
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Barchan, Karin
-
Herrlander, Dani ...
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Höjman, Patrik
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Karlsson, Marie
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Ljung, Lill
-
visa fler...
-
Andersson, Mats
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Bäckman, Eva
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Malmborg Hager, ...
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Walse, Björn
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Joosten, Leo
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van den Berg, Wi ...
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visa färre...
- Artiklar i publikationen
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Journal of Immun ...
- Av lärosätet
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Lunds universitet