Low-Cost Platform Technology for Rapid Isolation of Peripheral Blood Mononuclear Cells

Project Summary/Abstract CG Scientific aims to develop a low-cost, easy-to-use microfluidic device for rapid isolation of peripheral blood mononuclear cells (PBMCs), based on its patented platform technology named “High Efficiency Deterministic Separation (HEDS).” This Phase II project will focus on PBMCs because they are essential cells used in many areas of scientific and clinical research, including immunology, vaccine development, and drug discovery, as well as the study of auto-immune disorder, infectious disease, graft- versus-host disease, and cancer. The current standard method for PBMC isolation is Ficoll density gradient centrifugation, which is difficult to perform, requires substantial hands-on time, and is prone to user-to-user variations. Further, it provides sub-optimal cell recovery, introduces biases and phenotypic changes, and is inadequate for removing platelets. These drawbacks make the method unsuitable for many applications, especially those that require the results be consistent, accurate, and representative of disease states. To overcome these limitations, CG Scientific is developing an easy-to-use device that can isolate PBMCs from 10 ml of blood in 15 minutes, enabled by the HEDS technology. The technology has a unique configuration that allows for efficient PBMC isolation and low manufacturing cost — critical for the research market. In Phase I, the investigators have shown: (1) HEDS can greatly outperform Ficoll density gradient centrifugation and deliver outstanding ~97.2% PBMC recovery, ~99.97% RBC removal, and ~99.86% platelet depletion — with no loss of cell viability, no change in immunophenotype, and no T cell activation, (2) HEDS chips can be manufactured cost-effectively of cyclic olefin polymer (COP) using soft embossing, a mass production technique, and (3) the plastic prototype chip is functional, with ~82.3% PBMC recovery. This Phase II project aims to develop the technology all the way to a final, fully-validated prototype device, ready for pilot production, using 5 Specific Aims: (1) Finalize plastic chip design and manufacturing to provide >95% PBMC recovery, >99.8% RBC and platelet removal, and >40 ml/hr throughput; (2) Integrate the plastic chip into a 3D-printed cartridge to create an “alpha prototype” device that has the form factor and usability of the final product; (3) Develop a final, pre-production “beta prototype” using soft embossing and injection molding; (4) Characterize the cellular output of the beta prototype using flow cytometry, functional assays, and RNA sequencing; (5) Benchmark against standard Ficoll and immunomagnetic methods at multiple sites to establish the superiority of the HEDS device. The investigator team includes the inventor of HEDS, cell biologists, plastic manufacturing and medical device engineers, and directors of research and clinical laboratories that are regular users of Ficoll. The success of this project will lead to the development of an essential tool that will potentially become the standard for PBMC isolation, and make research in many fields more efficient, reproducible, and cost effective.