Rahimizadeh Parastou, Lee Jeong Eun, Lee Dae-Hee, In Lim Sung
Department of Chemical Engineering, Pukyong National University, 45 Yongso-ro, Namgu, Busan 48513, South Korea; Division of Research Center, Scripps Korea Antibody Institute, Chuncheon 24341, South Korea.
Department of Marine Food Science and Technology, Gangneung-Wonju National University, Gangneung, Gangwon-do 25457, South Korea; Nbios Inc, Gangneung, Gangwon-do 25457, South Korea.
Int J Biol Macromol. 2022 Apr 30;205:376-384. doi: 10.1016/j.ijbiomac.2022.02.040. Epub 2022 Feb 11.
A short in vivo half-life of protein-based therapeutics often restricts successful clinical translation despite their promising efficacy in vitro. As a biocompatible half-life extender, human serum albumin (HSA) has proven effective in some cases. While genetic fusion is well-established for interlinking HSA and a protein payload, it is limited to structurally simple proteins, necessitating new strategies to expand the utility of HSA for delivery of therapeutic proteins. Here, we report a novel HSA variant (eHSA) as a modular and long-acting carrier compatible with any protein payload of interest. The assembly between eHSA and a payload was driven by a heterodimeric coiled-coil interaction in which a short α-helix grafted onto HSA specifically bound to a complementary α-helix genetically fused to a payload. We showed various proteins including tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), single-chain TRAIL, or green fluorescent protein could piggyback onto eHSA via simple mixing without losing native activity. Additionally, either in presence or absence of a payload, eHSA was found to retain the pH-dependent FcRn-binding behavior - a critical attribute for prolonged survival in the systemic circulation. These results demonstrate eHSA would serve as a modular platform capable of delivering various therapeutic proteins with potentially long in vivo half-lives.
尽管基于蛋白质的治疗药物在体外显示出有前景的疗效，但其较短的体内半衰期常常限制了其成功的临床转化。作为一种生物相容性半衰期延长剂，人血清白蛋白（HSA）在某些情况下已被证明是有效的。虽然基因融合已被广泛用于连接HSA和蛋白质负载，但它仅限于结构简单的蛋白质，因此需要新的策略来扩大HSA在递送治疗性蛋白质方面的应用。在此，我们报道了一种新型的HSA变体（eHSA），它是一种模块化的长效载体，可与任何感兴趣的蛋白质负载兼容。eHSA与负载之间的组装由异源二聚体卷曲螺旋相互作用驱动，其中嫁接到HSA上的短α螺旋与基因融合到负载上的互补α螺旋特异性结合。我们表明，包括肿瘤坏死因子相关凋亡诱导配体（TRAIL）、单链TRAIL或绿色荧光蛋白在内的各种蛋白质可以通过简单混合搭载到eHSA上，而不会丧失天然活性。此外，无论是否存在负载，eHSA都被发现保留了pH依赖性的FcRn结合行为——这是在体循环中延长存活时间的关键属性。这些结果表明，eHSA将作为一个模块化平台，能够递送具有潜在长体内半衰期的各种治疗性蛋白质。
