A solid-phase PEGylation strategy for protein therapeutics using a potent FGF21 analog.
A solid-phase PEGylation strategy for protein therapeutics using a potent FGF21 analog.
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DOI：
10.1016/j.biomaterials.2014.03.023
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发表时间：
2014-06
期刊：
影响因子：
14
通讯作者：
Lintao Song;Yanlin Zhu;Huiyan Wang;A. A. Belov-A.;Jianlou Niu;Lu Shi;Yaoyao Xie;Chaohui Ye;
中科院分区：
文献类型：
--
作者：
Lintao Song;Yanlin Zhu;Huiyan Wang;A. A. Belov-A.;Jianlou Niu;Lu Shi;Yaoyao Xie;Chaohui Ye;
成纤维细胞生长因子21（FGF 21）是一种内分泌作用激素，具有治疗代谢性疾病的潜力，如2型糖尿病和肥胖症。由于FGF 21的低内在生物稳定性、聚集倾向及其对体内蛋白水解降解的敏感性，FGF 21发展为治疗剂受到阻碍。为了解决这些缺点，我们已经通过定点诱变策略性地引入半胱氨酸残基来开发重组人FGF 21变体，并且还开发了固相镍亲和PEG化策略，由此将固定化蛋白质的工程化的表面暴露的半胱氨酸残基用作与PEG-马来酰亚胺有效和位点选择性缀合的平台。工程化的PEG化FGF 21缀合物保留了其生物学功能，并且显示半衰期增加超过211.3分钟。通过证明FGF 21类似物作为原型的生物活性，我们还提供了有效增加蛋白质治疗剂的血清半衰期的“通用”固相方法。
Fibroblast growth factor 21 (FGF21) is an endocrine-acting hormone that has the potential to treat metabolic diseases, such as type 2 diabetes and obesity. Development of FGF21 into a therapeutic has been hindered due to its low intrinsic bio-stability, propensity towards aggregation and its susceptibility toin vivoproteolytic degradation. In order to address these shortcomings, we’ve developed recombinant human FGF21 variants by strategically introducing cysteine residues via site-directed mutagenesis, and have also developed a solid-phase nickel affinity PEGylation strategy, whereby engineered, surface-exposed cysteine residues of immobilized proteins were used as a platform to efficiently and site-selectively conjugate with PEG-maleimide. The engineered PEGylated FGF21 conjugates retained its biological functions, as well as demonstrated an increase in half-life by over 211.3 min. By demonstrating the biological activity of the FGF21 analog as a prototype, we have also provided a “generalized” solid-phase approach to effectively increase serum half-life of protein therapeutics.