SBIR Phase I: Continuous Near Infrared Monitor for Pichia Pastoris Bioreactors

该小型企业创新研究（SBIR）第一阶段项目将开发传感技术，能够在酵母毕赤酵母表达蛋白质期间连续监测关键代谢物的浓度。目前，连续监测是不可用的，并且使用耗时且劳动密集的离线分析来控制该过程。该传感技术使用连续收集的生物反应器生长培养基的近红外吸收光谱来监测分析物浓度和整个发酵过程中生物量的变化。初步结果表明，能够进行准确和连续的分析测量，并积极控制代谢物浓度。这一成功归功于ASL的创新，包括使用固态光谱仪，限制测量光谱范围，以及实施基于分析物特定化学信息的校准策略。第一阶段的工作将建立这种监测方法的商业可行性，通过开发一个强大的，实用的校准系统，用于常规操作，并通过设计一种手段，连续和非破坏性收集分析光谱，这项研究的更广泛的影响将是使生物反应器的准确反馈控制，从而加强优化工作，最大限度地提高高商品蛋白的生产产量。巴斯德毕赤酵母连续、实时监测系统的成功开发将增强其作为蛋白质表达平台的吸引力。拟议的监测器将克服离线分析的局限性，提供一种有效的手段，在发酵过程中，非破坏性地跟踪代谢物浓度和细胞密度的变化。这项传感技术将填补一般蛋白质表达市场的空白，在那里可靠的，在线传感是不可用的。该奖项是根据2009年美国复苏和再投资法案（公法111-5）资助。

This Small Business Innovation Research (SBIR) Phase I project will develop sensing technology capable of continuously monitoring concentrations of critical metabolites during protein expression with the yeast Pichia pastoris. Presently, continuous monitoring is unavailable and this process is controlled using time consuming and labor intensive off-line analyses. This sensing technology uses near infrared absorption spectra of bioreactor growth medium collected continuously to monitor analyte concentrations and changes in biomass throughout the fermentation process. Preliminary results demonstrate the ability to make accurate and continuous analytical measurements and to actively control metabolite concentrations. This success is attributed to ASL innovations that include using a solid-state spectrometer, restricting the measurement spectral range, and implementing a calibration strategy based on analyte-specific chemical information. This Phase I effort will establish the commercial feasibility of this monitoring approach by developing a robust, practical calibration system for routine operation and by designing a means to continuously and non-destructively collect analytical spectra.The broader impacts of this research will be to enable accurate feedback control of bioreactors, thereby enhancing optimization efforts and maximizing production yields of high commodity proteins. Successful development of a continuous, real-time monitor for Pichia pastoris will enhance its attractiveness as a protein expression platform. The proposed monitor will overcome the limitations of off-line analyses by providing an effective means to follow metabolite concentrations and cell density changes non-destructively during the course of a fermentation process. This sensing technology will fill a void in the general protein expression market where reliable, on-line sensing is unavailable.This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5).