Fusion of a short peptide that binds immunoglobulin G to a recombinant protein substantially increases its plasma half-life in mice.
Fusion of a short peptide that binds immunoglobulin G to a recombinant protein substantially increases its plasma half-life in mice.
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DOI：
10.1371/journal.pone.0102566
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Szoka FC
中科院分区：
文献类型：
--
作者：
Sockolosky JT;Kivimäe S;Szoka FC
我们探索了一种通过与Fc III基因融合来显著增加重组蛋白半衰期的策略，Fc III是一种13-mer IgG-Fc结构域结合肽（IgGBP），最初由DeLano及其同事在Genentech鉴定[DeLano WL，et al.（2000）Science 287∶1279-1283]。IgGBP融合通过使融合蛋白能够结合血清IgG来增加蛋白质的体内半衰期，这是最初由DeLano及其同事在专利中引入的概念，但据我们所知，这从未在科学文献中进行过。为了进一步研究IgGBP融合蛋白的体外和体内特性，我们将Fc III融合到模型荧光蛋白单体Katushka（mKate）的C-末端。mKate-IgGBP融合体易于在大肠杆菌中表达，并在pH 7.4和pH 6下分别以约40 nM和约20 nM的亲和力特异性结合人IgG，但不结合小鼠或大鼠IgG同种型。mKate-IgGBP在与人新生儿Fc受体（hFcRn）重叠的位点结合hIgG 1的Fc结构域，因此在体外抑制hIgG 1与hFcRn的结合。与人IgG的高亲和力结合还赋予mKate-IgGBP在小鼠中的长循环半衰期为108小时，与未修饰的mKate相比增加了75倍。因此，由于IgGBP的小尺寸，IgGBP融合通过搭载在血清IgG上而显著降低蛋白质清除率，而基本上不增加蛋白质分子量。这些有吸引力的特征可能导致蛋白质疗法具有降低的剂量频率和改善的患者依从性。
We explore a strategy to substantially increase the half-life of recombinant proteins by genetic fusion to FcIII, a 13-mer IgG-Fc domain binding peptide (IgGBP) originally identified by DeLano and co-workers at Genentech [DeLano WL, et al. (2000) Science 287∶1279–1283]. IgGBP fusion increases the in vivo half-life of proteins by enabling the fusion protein to bind serum IgG, a concept originally introduced by DeLano and co-workers in a patent but that to the best of our knowledge has never been pursued in the scientific literature. To further investigate the in vitro and in vivo properties of IgGBP fusion proteins, we fused FcIII to the C-terminus of a model fluorescent protein, monomeric Katushka (mKate). mKate-IgGBP fusions are easily expressed in Escherichia coli and bind specifically to human IgG with an affinity of ∼40 nM and ∼20 nM at pH 7.4 and pH 6, respectively, but not to mouse or rat IgG isotypes. mKate-IgGBP binds the Fc-domain of hIgG1 at a site overlapping the human neonatal Fc receptor (hFcRn) and as a consequence inhibits the binding of hIgG1 to hFcRn in vitro. High affinity binding to human IgG also endows mKate-IgGBP with a long circulation half-life of ∼8 hr in mice, a 75-fold increase compared to unmodified mKate. Thus, IgGBP fusion significantly reduces protein clearance by piggybacking on serum IgG without substantially increasing protein molecular weight due to the small size of the IgGBP. These attractive features could result in protein therapies with reduced dose frequency and improved patient compliance.
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影响因子：
9
作者：
Rath T;Baker K;Dumont JA;Peters RT;Jiang H;Qiao SW;Lencer WI;Pierce GF;Blumberg RS
通讯作者：
Blumberg RS
影响因子：
56.9
作者：
DeLano, WL;Ultsch, MH;Wells, JA
通讯作者：
Wells, JA
影响因子：
2.4
作者：
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通讯作者：
Tomlinson, Ian M.
影响因子：
4.4
作者：
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通讯作者：
Roopenian, Derry C.
影响因子：
4.4
作者：
Ober, RJ;Radu, CG;Ward, ES
通讯作者：
Ward, ES