3D-Printed Tangential Flow Filtration and High-Throughput Microfluidic Electroporation for Scalable Microbial Processing

Abstract

Bacterial transformation via electroporation is fundamental to modern biotechnology applications including therapeutic protein production, biomaterial synthesis, and agricultural enhancement. However, conventional electroporation workflows face critical bottlenecks that limit their scalability and industrial applicability; mainly inefficient electrocompetent cell preparation and low-throughput transformation processes. This thesis presents two complementary 3D-printed technologies that independently address these limitations for scalable microbial processing. First, a novel spiral channel tangential flow filtration (TFF) system was developed that replaces conventional centrifugation-based methods for preparing electrocompetent cells. The spiral geometry enhances mixing dynamics and enables continuous washing of bacterial cultures, dramatically reducing preparation time while improving cell recovery compared to traditional centrifugation and membrane filtration approaches that suffer from time constraints, labor intensity, and membrane fouling. Second, a 3D-printed microfluidic electroporation platform featuring geometry-optimized electric field distribution was designed. Building upon established M-TUBE principles, the bilaterally converged channel architecture creates localized field enhancement at reduced applied voltages, enabling high-efficiency transformation of larger cell volumes. This design overcomes the throughput limitations of conventional cuvette-based systems that require manual handling and process only small volumes. Both technologies leverage additive manufacturing to create cost-effective alternatives to traditional protocols. Computational fluid dynamics simulations and experimental validation demonstrate significant improvements in processing time, transformation efficiency, and throughput compared to conventional methods. These complementary technologies demonstrate the potential for future integration into a complete workflow for scalable microbial transformation, with promising implications for broader implementation in industrial biotechnology, synthetic biology, and large-scale research applications.