PFI-TT: A High-Throughput Instrument for Measuring the Stability of Liquid Formulations of Therapeutic Proteins

这一创新-技术转化伙伴关系(PFI-TT)项目的更广泛影响/商业潜力是开发一种仪器，它将减少筛选治疗性蛋白质稳定性所需的时间，治疗性蛋白质已经彻底改变了抗击癌症、阿尔茨海默氏症和关节炎等疾病的能力。这些以蛋白质为基础的药物的稳定性是确保液体制剂在冷藏温度下保质期为2年的主要问题。目前，行业评估标准包括在压力储存条件下进行为期1个月的稳定性试验。提高替代前配方和配方筛选工具的效率和准确性应该会减少药物开发第一阶段所需的时间，并使探索更多的蛋白质和更多的溶液条件成为可能。预计将这项技术纳入配方工作流程将有助于增加可推向市场的治疗性蛋白质的数量，并降低销售产品的成本。拟议的项目涉及建造一种设备，旨在测量治疗性蛋白质对相分离的敏感度。像油和水一样，浓缩的治疗性蛋白质溶液在相分离温度以下可以分成两个不同的相，在那里溶液变得混浊。相分离温度和相分离动力学为不稳定性提供了有价值的量度，并且可以通过暗场显微镜的简单光散射测量来监测。在这里，我们概述了温度梯度微流控装置的硬件和软件的开发，该装置可以诱导相分离并提取描述相变潜在的蛋白质-蛋白质相互作用的参数。该项目的目标是设计一个用于控制温度梯度、光学仪器和数据分析的自动化系统。最终，这些研究和开发工作将提供温度梯度微流控技术的原型版本，该技术可用于测量各种治疗性蛋白质的稳定性。这种高通量测量将能够探索导致配方不稳定性的pH、盐、缓冲液和蛋白质化学。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this Partnerships for Innovation-Technology Translation (PFI-TT) project is the development of an instrument that will reduce the time required to screen the stability of therapeutic proteins, which have revolutionized the ability to fight diseases like cancer, Alzheimer's disease, and arthritis. The stability of these protein-based drugs is a major concern for ensuring a 2-year shelf-life for liquid formulations under refrigerated temperatures. Currently, the industry assessment standard involves 1-month long stability tests under stressed storage conditions. Improving the efficiency and accuracy of alternative pre-formulation and formulation screening tools should reduce the time required for phase 1 of drug development and make it possible to explore more proteins and more solution conditions. It is expected that the incorporation of this technology into the formulation workflow should help increase the number of therapeutic proteins that can be brought to market and reduce the cost of the products being sold. The proposed project involves the construction of a device designed to measure the susceptibility of a therapeutic protein to phase separation. Like oil and water, concentrated therapeutic protein solutions can separate into two distinct phases below a phase separation temperature, where the solution becomes cloudy. The phase separation temperature and the kinetics of the phase separation provide valuable metrics for instability and can be monitored by simple light scattering measurements via dark-field microscopy. Herein, we outline the development of hardware and software for a temperature gradient microfluidics device that can induce phase separation and extract parameters describing the underlying protein-protein interactions of the phase transition. The goals of this project are to engineer an automated system for controlling the temperature gradient, optical instrumentation, and data analysis. Ultimately, these research and development efforts will provide a prototype version of the temperature gradient microfluidics technology that can be implemented for measuring the stability of a variety of therapeutic proteins. Such high-throughput measurements will enable the exploration of pH, salt, buffers, and protein chemistries that contribute to formulation instability.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.