期刊
ACS BIOMATERIALS SCIENCE & ENGINEERING
卷 3, 期 3, 页码 304-312出版社
AMER CHEMICAL SOC
DOI: 10.1021/acsbiomaterials.6b00578
关键词
cytokine engineering; 2-methyl-2-oxazoline; genetic code expansion; CuAAC (copper(I) catalyzed azide alkyne cycloaddition); bioconjugation
资金
- DFG [ME 3920/3-1]
- Sino-German center
- Bundesministerium fur Bildung und Forschung (Germany) [13N13454, 081220/127]
- Carl-Zeiss Foundation (JCSM Strukturantrag)
- Thuringer Ministerium fur Wirtschaft, Wissenschaft, und Digitale Gesellschaft (TMWWDG, Pro-Exzellenz II, NanoPolar)
- German Research Foundation (DFG) [GZ: HA 7725/1-1]
Polymer conjugated biologics form a multibillion dollar market, dominated by poly(ethylene glycol) (PEG). Recent reports linked PEGs to immunological concerns, fueling the need for alternative polymers. Therefore, we are presenting a strategy replacing PEG by poly(2-oxazoline) (POx) polymers using genetically engineered interleukin-4 (IL-4) featuring an unnatural amino acid for site-specific conjugation through bioorthogonal copper-catalyzed azide alkyne cycloaddition (CuAAC). Conjugation yields of 1L-4-PEG were poor and did not respond to an increase in the copper catalyst. In contrast, POxylated IL-4 conjugates resulted in homogeneous conjugate outcome, as demonstrated electrophoretically by size exclusion chromatography and analytical ultracentrifugation. Furthermore, POxylation did not impair thermal and chemical stability, and preserved wild-type IL-4 activity for the conjugates as demonstrated by TF-1 cell proliferation and STAT-6 phosphorylation in HEK293T cells, respectively. In conclusion, POxylation provides an interesting alternative to PEGylation with superior outcome for the synthesis yield by CuAAC and resulting in conjugates with excellent thermal and chemical stress profiles as well as biological performances.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据