Commercial Development of a Novel Microfluidic Platform for Real-Time Biopharmaceutical Analysis

项目摘要 -- 生物药物是在活细胞中制造的复杂分子，因此经常 质量测试是开发、监管批准和制造所必需的。毛细管等电点 聚焦(CIEF)是一种广泛使用的分析技术，可以快速检测大多数蛋白质 修改。质谱仪(MS)测量固有分子量，并具有独特的能力 确定特定蛋白质修饰的特定性质。CIEF和MS的结合是一个强大和高度 确定蛋白质异构体之间组成差异的有效方法。但是，当前 方法学需要劳动密集型制备层析法来分离足够的物质 结合CIEF和MS分析，因此在容量和吞吐量方面受到严重限制。为了解决这个问题， Intabio已经开发了Blaze™系统，这是第一个拥有专利的蛋白质分析平台 商业上可用的系统，将CIEF与MS分析无缝集成。Blaze系统利用一种 集成(1)等电聚焦分离蛋白质亚型，(2)成像的新型微流控芯片设计 通过280 nm吸收测定蛋白质异构体，以及(3)MS样品制备和 通过电喷雾进入相邻的质谱计以检测和鉴定每一种异构体。大火 系统集成了这些步骤，大大减少了分析时间，每个样品只需几分钟。 这一SBIR项目将推动BLAZE仪器、软件和消耗品套件的开发， 并将建立消耗品制造流程，为商业投放做准备。第一阶段将专注于 关于推进核心创新-芯片和翻盖。翻盖将微流控芯片固定在 仪器作为世界到芯片的接口，提供试剂、样品、压力和 电极。在第一阶段，将优化三层芯片的激光键合工艺，以提高成品率和 芯片性能通过探索多种键合参数组合来实现。此外，原型设计还可以 为了提高易用性，将把芯片和翻盖集成到一个单独的“墨盒”中，将进行设计和测试。 第二阶段将把原型墨盒推进到商业、一次性版本，并设置商业 芯片盒的制造工艺。商业仪器和软件开发也将 在第二阶段完成。仪器开发将解决降低材料成本的战略，并将 内部自动取样器功能；软件开发将完善CIEF和MS数据的集成和显示，如 以及完全符合cfr21part11。 该项目的创新和研究将推动蛋白质分离科学和技术领域的发展 拓展微流控技术领域。该项目还将改变生物制药药物的开发。 并通过实现更频繁、更全面的测试来实现制造。这将带来极大的生产力。 收益，转化为以更短的时间和更低的成本开发出更多更高质量的药物。

Project Summary   Biopharmaceuticals are complex molecules manufactured in living cells, and as a result, frequent quality testing is required for development, regulatory approval, and manufacturing. Capillary isoelectric focusing (cIEF) is a widely used analytical technique that provides rapid detection of most protein modifications. Mass spectrometry (MS) measures intrinsic molecular weight, and has the unique capability of identifying the specific nature of a given protein modification. Combining cIEF and MS is a powerful and highly effective approach to determining compositional differences between protein isoforms. However, current methodologies require labor intensive preparative chromatography to isolate sufficient material for the combined cIEF and MS analysis, and so are severely limited in capacity and throughput. To address this issue, Intabio has developed the Blaze™ system, a proprietary protein analytics platform that will be the first commercially available system to seamlessly integrate cIEF with MS analysis. The Blaze system utilizes a novel microfluidic chip design to integrate (1) separation of protein isoforms by isoelectric focusing, (2) imaging of protein isoforms by 280nm absorption for detection and quantitation, and (3) MS sample preparation and delivery by electrospray into an adjacent mass spectrometer to detect and identify each isoform. The Blaze system's integration of these steps drastically reduces the analysis time to only minutes per sample. This SBIR project will advance the development of the Blaze instrument, software and consumable kit, and will establish a consumable manufacturing process to prepare for commercial launch. Phase 1 will focus on advancing the core innovation - the chip and clamshell. The clamshell holds the microfluidic chip in the instrument and acts as the world-to-chip-interface, providing inputs for reagents, samples, pressure and electrodes. In Phase 1, the laser bonding process for the three-layer chip will be optimized to improve yield and chip performance by exploring multiple combinations of bonding parameters. Additionally, prototype designs to integrate the chip and clamshell into a single “cartridge” for improved ease-of-use will be designed and tested. Phase 2 will advance the prototype cartridge to a commercial, disposable version and set up the commercial manufacturing process for the chip cartridge. Commercial instrument and software development will also be completed in Phase 2. Instrument development will address strategies to reduce material cost and integrate an internal autosampler function; software development will refine integration and display of cIEF and MS data, as well as complete cfr21part11 compliance. The innovation and research from this project will advance the fields of protein separation science and expand the microfluidic technology field. This project will also transform biopharmaceutical drug development and manufacturing by enabling more frequent, comprehensive testing. This will result in profound productivity gains, translating into a greater number of higher quality drugs developed in less time and with less cost.