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

项目摘要 生物制药是在活细胞中制造的复杂分子，因此， 开发、监管批准和制造都需要质量测试。毛细管等电 聚焦（cIEF）是一种广泛使用的分析技术，可快速检测大多数蛋白质 修改.质谱法（MS）测量固有分子量，并且具有独特的能力， 鉴定给定蛋白质修饰的特定性质。结合cIEF和MS是一个强大的，高度 确定蛋白质异构体之间的组成差异的有效方法。但目前的 这些方法需要劳动密集型制备色谱法来分离足够的材料， cIEF和MS分析的组合，因此在容量和通量方面受到严重限制。为了解决这个问题， Intabio开发了Blaze ™系统，这是一个专有的蛋白质分析平台，将成为第一个 将cIEF与MS分析无缝集成。Blaze系统利用 新型微流控芯片设计，其集成（1）通过等电聚焦分离蛋白质同种型，（2）成像 通过280 nm吸收测定蛋白质同种型，用于检测和定量，以及（3）MS样品制备和 通过电喷雾递送到相邻的质谱仪中以检测和鉴定每种同种型。大火 系统集成了这些步骤，大大缩短了每个样品的分析时间，仅需几分钟。 该SBIR项目将推进Blaze仪器、软件和耗材套件的开发， 并将建立一个消耗品制造过程，为商业发射做准备。第一阶段将重点 推进核心创新-芯片和翻盖。蛤壳将微流体芯片保持在 仪器，并作为世界到芯片的接口，提供试剂，样品，压力和 个电极在第一阶段，将优化三层芯片的激光键合工艺，以提高产量， 通过探索键合参数的多种组合来提高芯片性能。此外，原型设计 将设计和测试将芯片和翻盖集成到一个单一的"墨盒"，以提高易用性。 第2阶段将把原型盒推进到商业化的一次性版本，并建立商业化的 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和集成 内部自动进样器功能;软件开发将完善cIEF和MS数据的整合和显示， 以及完全符合《联邦财务条例》第21部分第11条。 该项目的创新和研究将推动蛋白质分离科学领域的发展， 拓展微流控技术领域。该项目还将改变生物制药药物开发 和制造，实现更频繁、更全面的测试。这将带来深远的生产力 这些成果转化为在更短的时间内以更低的成本开发出更多高质量的药物。

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.