Continuous Countercurrent Tangential Chromatography

描述(由申请人提供)：国家卫生研究院的SBIR第一阶段提案要求为Chromatan，Inc.提供183,000美元的资金，以证明使用连续逆流切向色谱(CCTC)纯化高价值生物制品的可行性，例如用于治疗癌症的单抗或用于预防病毒疾病或流感大流行的重组疫苗。下游加工目前占重组蛋白产品生产总成本的80%之多。由于填充床色谱柱的容量有限和费用高昂，细胞培养技术的最新进展已经造成了“下游瓶颈”。CCTC克服了传统柱层析的许多局限性，它使用浆液形式的树脂颗粒，通过由一系列静态混合器和中空纤维膜组件组成的一次性流动通道泵送。渗透液的逆流循环被用来显著提高工艺效率并减少树脂和缓冲液的需求，类似于逆流液-液萃取的行为。CCTC的最佳膜孔径、树脂粒径和操作条件(如交叉流速、渗透流速和在静态混合器中的停留时间)将通过对临界滤液通量和结合/洗脱动力学的实验测量来确定。CCTC可以使用比填充床柱小得多的树脂颗粒，因为不再有任何压降限制，这可以提供显著更快的传质速率。这项研究的总体目标是设计、建造和测试一个原型CCTC系统，该系统能够以大于90%的产品产率和95%的纯度分离模型蛋白质混合物。Chromatan预计，CCTC技术将为层析的大规模商业应用提供以下有利影响：与柱层析相比，树脂体积需求减少5倍；一次性流路，无需进行昂贵的清洗验证；可直接扩展到20,000批L；大规模操作中的资本成本显著降低(高达60%)；捕获层析步骤的总体成本显著节省(高达75%)。这项技术的持续性和一次性性质将通过缩短上市时间和降低生产成本来改善公众健康，这两者都将导致药品成本的降低。Chromatan预计，第五年的市场销售额将达到10%，收入将达到1亿美元。 公共卫生相关性：拟议的SBIR项目的总体目标是设计、建造和测试一种能够提供高效、低成本的治疗性蛋白质和疫苗纯化的连续逆流切向层析系统。与当前的分离技术相比，CCTC的设计具有一次性流程、连续运行、较低的资本和运营成本，并将通过降低重要药物产品的成本和减少将新的治疗药物推向市场所需的时间，显著改善公众健康。

DESCRIPTION (provided by applicant): SBIR Phase I proposal to NIH requests $183,000 funding for Chromatan, Inc. to demonstrate the feasibility of using Continuous Countercurrent Tangential Chromatography (CCTC) for the purification of high value biological products, e.g., monoclonal antibodies for the treatment of cancer or recombinant vaccines for protection against viral disease or a flu pandemic. Downstream processing currently accounts for as much as 80% of the overall cost of production of recombinant protein products. Recent advances in cell culture technology have created a "downstream bottleneck" due to the limited capacity and high expense of packed bed chromatography columns. CCTC overcomes many of the limitations of conventional column chromatography by using resin particles in the form of a slurry, which is pumped through a disposable flow path consisting of a series of static mixers and hollow fiber membrane modules. Countercurrent recycling of the permeate is used to significantly increase process efficiency and reduce resin and buffer requirements, analogous to the behavior in countercurrent liquid-liquid extraction. The optimal membrane pore size, resin particle size, and operating conditions for CCTC (e.g. crossflow rate, permeate flow rate, and residence time in the static mixers) will be determined from experimental measurements of the critical filtrate flux and the binding / elution kinetics. CCTC can employ much smaller resin particles than packed bed columns, since there is no longer any pressure drop limitation, which can provide significantly faster mass transfer rates. The overall goal of the proposed research is to design, construct, and test a prototype CCTC system capable of separating a model protein mixture with greater than 90% product yield and 95% purity. Chromatan envisions that CCTC technology will provide the following favorable impacts for large-scale commercial applications of chromatography: 5-fold reduction in resin volume requirements compared with column chromatography; a disposable flow path that eliminates the need for costly cleaning validation; straightforward scalability up to batch volumes of 20,000 L; significant reduction of capital costs (up to 60%) in large-scale operations; significant overall cost savings (up to 75%) for capture chromatography steps. The continuous and disposable nature of this technology will improve public health by decreasing time to market and reducing production costs, both of which will lead to a reduction in drug costs. Chromatan projects a fifth year market peneratraion of 10% and revenues of $100 million. PUBLIC HEALTH RELEVANCE: The overall goal of the proposed SBIR project is to design, construct, and test a Continuous Countercurrent Tangential Chromatography system capable of providing highly efficient and low cost purification of therapeutic proteins and vaccines. CCTC is designed to have a disposable flow path, continuous operation, and lower capital and operating costs compared to current separation technologies, and will lead to significant improvements in public health by reducing the cost of important drug products, and by decreasing the time required to bring new therapeutic drugs to market.