Stability of a high-concentration monoclonal antibody solution produced by liquid-liquid phase separation.
Stability of a high-concentration monoclonal antibody solution produced by liquid-liquid phase separation.
复制标题
DOI：
10.1080/19420862.2021.1940666
复制
发表时间：
2021-01
期刊：
影响因子：
5.3
通讯作者：
Golovanov AP
中科院分区：
文献类型：
--
作者：
Bramham JE;Davies SA;Podmore A;Golovanov AP
皮下注射低体积（<2 mL）高浓度（>100 mg/mL）制剂是单克隆抗体（mAb）和其他生物制药蛋白质的有吸引力的给药策略。使用浓缩溶液在生物加工的各个阶段也可能是有益的。然而，通过常规技术如超滤浓缩蛋白质可能是耗时且具有挑战性的。通过宏观液-液相分离（LLPS）产生的致密级分的分离已经被建议作为产生高浓度溶液的手段，但是该方法的实用性和所得蛋白质溶液的稳定性先前尚未被证明。在这项概念验证研究中，我们证明LLPS可用于将mAb溶液浓缩至>170 mg/mL。我们表明，单克隆抗体的结构是不改变LLPS，和未扰动的单克隆抗体是可回收的稠密部分稀释后，判断由1H核磁共振光谱。最后，我们表明，从致密部分获得的模型高浓度蛋白质制剂的物理性质和稳定性可以得到改善，例如通过添加赋形剂精氨酸·谷氨酸盐。这导致稳定的高浓度蛋白质制剂具有降低的粘度并且没有进一步的宏观LLPS。通过LLPS浓缩mAb溶液代表了一种简单有效的技术，可用于生产用于生物加工或给药的高浓度蛋白质制剂。缩略语精氨酸·谷氨酸（Arg·Glu）、Carr-Purcell-Meiboom-Gill（CPMG）、临界温度（TC）、高效分子排阻色谱（HPSEC）、液-液相分离（LLPS）、单克隆抗体（mAb）、核磁共振（NMR）、横向弛豫速率（R2）
Subcutaneous injection of a low volume (<2 mL) high concentration (>100 mg/mL) formulation is an attractive administration strategy for monoclonal antibodies (mAbs) and other biopharmaceutical proteins. Using concentrated solutions may also be beneficial at various stages of bioprocessing. However, concentrating proteins by conventional techniques, such as ultrafiltration, can be time consuming and challenging. Isolation of the dense fraction produced by macroscopic liquid–liquid phase separation (LLPS) has been suggested as a means to produce high-concentration solutions, but practicality of this method, and the stability of the resulting protein solution have not previously been demonstrated. In this proof-of-concept study, we demonstrate that LLPS can be used to concentrate a mAb solution to >170 mg/mL. We show that the structure of the mAb is not altered by LLPS, and unperturbed mAb is recoverable following dilution of the dense fraction, as judged by 1H nuclear magnetic resonance spectroscopy. Finally, we show that the physical properties and stability of a model high concentration protein formulation obtained from the dense fraction can be improved, for example through the addition of the excipient arginine·glutamate. This results in a stable high-concentration protein formulation with reduced viscosity and no further macroscopic LLPS. Concentrating mAb solutions by LLPS represents a simple and effective technique to progress toward producing high-concentration protein formulations for bioprocessing or administration. Abbreviations Arginine·glutamate (Arg·Glu), Carr-Purcell-Meiboom-Gill (CPMG), critical temperature (TC), high-performance size-exclusion chromatography (HPSEC), liquid–liquid phase separation (LLPS), monoclonal antibody (mAb), nuclear magnetic resonance (NMR), transverse relaxation rate (R2)
登录
查看更多内容
影响因子：
--
作者：
Bramham JE;Podmore A;Davies SA;Golovanov AP
通讯作者：
Golovanov AP
影响因子：
3.8
作者：
Allmendinger, Andrea;Mueller, Robert;Fischer, Stefan
通讯作者：
Fischer, Stefan
影响因子：
4.8
作者：
Joubert, Marisa K.;Luo, Quanzhou;Narhi, Linda O.
通讯作者：
Narhi, Linda O.
影响因子：
5.3
作者：
Kheddo P;Cliff MJ;Uddin S;van der Walle CF;Golovanov AP
通讯作者：
Golovanov AP
影响因子：
3.7
作者：
Nishi, Hirotaka;Miyajima, Makoto;Fukui, Kiichi
通讯作者：
Fukui, Kiichi