Miniaturisation of high throughput healthcare bioreactors for advanced therapeutics

鉴于诺华公司Kymriah免疫疗法的成功，利用细胞和基因疗法（CDGT）对患者自身细胞进行重新编程以对抗退行性疾病的前景毫无疑问。然而，一个主要问题仍然存在：价格点。CGT的巨大成本反映了当前的技术水平：从发现到商业化的制造和开发过程缓慢，同时执行不同工艺所需的多个设备的成本过高。因此，瓶颈存在于生产和可访问性之间。目前，对一个强大的、可扩展的和封闭的生物加工制造平台的需求尚未得到满足，该平台可以实现安全、低成本的治疗，并缩短开发时间，以提高患者的可及性。我们的主要目标是开发一个基于微流体技术的高通量生物处理平台。微流体-在微尺度流体操作的科学-能够解决流线型和高通量细胞培养生产的挑战，通过优化细胞扩增过程中的流体消耗。我们已经提交了微流控芯片设计的欧洲专利申请（B74637EP D38585），这是生物处理平台设备的核心。这种设计不同于现有的微流体工作，因为它是多功能的，即能够在密封环境中原位执行标准过程（播种、转导、洗涤、取样、收获），同时防止侵入性干预。与传统的2D/3D制造相比，我们的创新来自于应用于大规模细胞培养的微流体技术。与传统方法相比，微流控细胞培养技术具有以下优点：能够在封闭系统中进行播种，扩增，转导，分化，过滤，采样和收获过程。传统的2D/3D系统，甚至目前的小规模微流体系统都具有有限的功能（即设想执行一种或几种功能，例如膨胀和灌注），因此需要在某些时候“打开”过程，这通常是劳动密集型的（因此成本高昂），并构成安全威胁（例如污染）。2. 显著减少反应物消耗：由于固有的微小体积加上微流体中使用的连续灌注系统，通常降低反应物消耗10-20倍。目前，使用传统的2D/3D系统，反应物占总成本的30-35%。更好地控制工艺参数。2D/3D系统的特点是电池与总体积比高，导致最终产品不均匀，工艺效率低（例如，转导电池需要1-2天）。微流体可以精细地控制浓度并最大限度地提高细胞与反应物的接触，即由于微通道中的流体循环，细胞可以平等地获得氧气和营养物质，从而产生更均匀的最终产品和更少的过程失败（例如细胞死亡）。4. 能够在不改变流程的情况下扩大规模。传统2D/3D技术的放大需要工艺调整。现有的小型微流体系统不是为高通量设计的，因此不能以经济有效的方式进行扩展。我们的平台使用可堆叠的盒式系统，可以从几百个单元扩展到数亿个单元，无需任何工艺调整。细胞和流体可以通过自动泵和微流体阀在系统的不同部分之间转移。响应这些规范的生物处理平台的发明需要多学科方法和对基本科学原理的理解：流体动力学（流体力学），细胞生长和培养（生物学/生物物理学/生物化学），以及用于并行化和自动化多个大型芯片的硬件（工程）的开发。

There is little doubt of the promise of reprogramming a patient's own cells to combat degenerative diseases using cell and gene therapies (CDGT), given the success of Novartis Kymriah immunotherapy. However, one principal problem persists: the price point. The colossal costs of CGT reflect the current state-of-the-art: a slow manufacturing and development process from discovery to commercialization along with the exorbitant costs of multiple equipment needed to perform different processes. Thus, the bottleneck exists between production and accessibility. There is currently an unfulfilled need for a robust, scalable and closed bioprocessing manufacturing platform that can enable safe, low cost treatments, and rapid development time to improve accessibility to patients.Our primary goal is to develop a high-throughput bioprocessing platform based on microfluidics technology. Microfluidics - the science of fluid manipulation in the microscale - is able to address the challenges for streamlined and high-throughput cell culture production by optimizing fluid consumption during cell expansion. We have already submitted a European patent application (B74637EP D38585) of a microfluidic chip design, which is the heart of the bioprocessing platform device. This design differs from existing microfluidic work because it is multi-functional, i.e. capable of performing standard processes (seeding, transduction, washing, sampling, harvest) in situ in a sealed environment while preventing invasive interventions.Our innovation comes from the microfluidic technology applied to large scale cell culture compared to conventional 2D/3D manufacturing. The microfluidic cell culture technology has demonstrated several advantages compared to conventional methods:1. Ability to perform seeding, expansion, transduction, differentiation, filtration, sampling and harvest processes in a closed system. Conventional 2D/3D systems and even current small-scale microfluidic systems have limited functionality (i.e. conceived to perform one or a few functions, e.g. expansion and perfusion) and therefore require 'opening' the process at some point, which usually is labor-intensive (therefore costly) and poses a safety threat (e.g. contamination). 2. Dramatic reduction in reactant consumption: usually 10-20x lower reactant consumption due to inherent minute volumes coupled with continuous perfusion systems used in microfluidics. Currently, with conventional 2D/3D systems, reactants represent 30-35% of total costs.3. Better control of process parameters. 2D/3D systems are characterized by high cell to total volume ratio which leads to heterogeneous end product and low process efficiency (e.g., 1-2 days to transduce cells). Microfluidics allows to finely control concentrations and maximize cell-to-reactant contact, i.e. cells have equal access to oxygen and nutrients due to fluid circulation in micro-channels, which results in a more homogeneous end product and less process failure (e.g. cell death). 4. Ability to scale up without process change. Scale-ups with conventional 2D/3D technology require process adaptation. Existing small-scale microfluidic systems are not designed for high-throughput and thus cannot be scaled in a cost-effective way. Our platform uses a stackable cassette system which can be scaled from a few hundred cells to several hundred million of cells, without any process adaptations. Cells and fluids can be transferred between different parts of the system via automated pumps and microfluidic valves.The invention of a bioprocessing platform that responds to these specifications requires multi-disciplinary approach and understanding of the underlying scientific principles: flow hydrodynamics (fluid mechanics), cell growth and culture (biology/biophysics/biochemistry), together with the development of the hardware (engineering) used for parallelization and automation of multiple large chips.