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在接触式流量过过程中,内部模块压力概况和蛋白质积累

Fernanda Cunha^1,2, Jessica Zuponcic^1,2, Francesco Rossi³

¹Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, Indiana, USA.

Biotechnology progress

|September 25, 2023

概括

接触式流量过 (TFF) 在高蛋白度下面面临流量下降. 这项研究测量了蛋白质沉积和压力下降,揭示了50微米层限制了单克隆抗体 (mAb) 净化中的超过率.

科学领域:

生物技术是生物技术.
化学工程是化学工程的重要组成部分.
膜科学膜科学膜科学

背景情况:

使用30kDa超膜的接触式流过 (TFF) 是缩单克隆抗体 (mAbs) 的标准.
在TFF过程中蛋白质度高导致过率 (流量) 显著下降,阻碍生物制药制造.
单克隆抗体 (mAbs),通常是IgG类型蛋白质 (140-160kDa),需要高度 (≥200 mg/mL) 的配方.

研究的目的:

直接测量高度蛋白质TFF期间的局部跨膜压力下降.
使用共聚焦成像,可视化膜表面上的蛋白质积累.
了解在mAb净化过程中限制超率的机制.

主要方法:

使用修改后的膜系统直接测量局部的跨膜压力下降.
采用离线共聚焦激光扫描显微镜来图像蛋白质积累.
在超过200 mg/mL的度下使用150 kDa的牛IgG进行实验.

主要成果:

沿膜长度测量了大约1.2 psi/cm的恒定跨膜压力下降.
在高蛋白度下,在膜表面观察到一个50微米厚的均沉积的蛋白质层.
证实轴向压力在膜磁带上下降了58psi.

结论:

蛋白质沉积是限制缩mAbs.TFF中的超过率的主要因素.
局部测量为膜性能限制提供了关键的见解.
结果表明,在生物制药制造中改善TFF膜性能的潜在目标.

关键词:

与焦点一致的激光扫描显微镜 (CLSM)凝层是一种凝层.这是一种单克隆抗体.蛋白质沉积是一种蛋白质沉积.接触式流量过的过器.

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