SBIR-STTR Award

The goal of this Fast-Track STTR project is to develop a Deterministic Lateral Displacement (DLD) microfluidic device that can enrich white blood cells (WBCs) from a typical leukapheresis unit in 1 hr, for use in manufacturing cancer cellular immunotherapy. Chimeric antigen receptor T cell (CAR-T) therapy has been recommended for FDA approval to treat relapsed or refractory pediatric and young adult patients with B-cell acute lymphoblastic leukemia. There is a critical need for cost-effective automated methods to improve the efficiency and yield of large-scale enrichment of WBCs for use in manufacturing CAR-T and other cellular therapies. GPB is a pioneer in developing novel DLD microchips to process blood cells for cell analysis (19,26). GPB now proposes to develop, evaluate and commercialize a compact device in which an entire leukapheresis unit (up to 5x1010 WBCs in up to 300 ml) can be processed in a ?Leuko-stack? of disposable single-use multi-channel DLD chips to produce in 1 hr a washed cell suspension that is enriched in WBCs and depleted of red blood cells (RBCs) and platelets (PLT). In Phase I, Aim 1 is to increase cell throughput through the current prototype chips by: 1) optimizing DLD chip design and operation to increase flow rate; 2) increasing throughput by stacking plastic chips and running them in parallel (?Leuko-stacks?); and 3) translating chip production to high-volume manufacturing material such as Cyclic Olefin Polymer (COP). Final Phase I milestones to proceed to Phase II are: 1) final chip design with a flow rate of at least 25 mL/hr via a single chip, at least 70% recovery of viable WBCs and immunophenotype- defined T-lymphocytes, and ability to process cells for 1 hr without clogging; 2) Leuko-stack of at least 6 chips run in parallel, with the same output as in #1; 3) combined increases in throughput via #1 and #2 sufficient to process a 300 ml leukapheresis unit in 1 hr; 4) confirmation that the chips can be produced from COP. In Phase II, Aim 2 is to build final prototype COP plastic chip-based microfluidic device capable of processing a leukapheresis sample at 300 mL/hr. Aim 3 is to test performance of prototypes from Aim 2 with leukapheresis aliquots and then full-size human leukapheresis samples. The final milestone of this project is to produce a set of commercial prototype Leuko-stacks that can process an entire 300-ml leukapheresis unit in 1 hr with at least 70% WBC and T-lymphocyte recovery, at least 90% depletion of RBCs, at least 80% depletion of PLTs, and at least 70% recovery of T-cell expansion capacity (as compared with the input samples) in significantly more than 50% of samples tested at 2 sites. The GPB Leuko-stack platform will preserve the advantages of DLD microfluidic cell processing over current methods, while massively increasing throughput rate and cell processing capacity, thus transitioning from analytic- to preparative-scale WBC enrichment for subsequent manufacture of CAR-T and other cell therapies.

Project Terms:
Aliquot; anticancer research; Autologous; B-Cell Acute Leukemia; base; Blood; Blood Cells; Blood Platelets; Businesses; cancer cell; cell injury; Cell physiology; Cell Separation; Cell Therapy; cell type; Cells; Cellular immunotherapy; cellular targeting; Centrifugation; Childhood; chimeric antigen receptor; Clinical; Clinical Trials; Collaborations; cost; cost effective; Cycloparaffins; design; Device or Instrument Development; Devices; Doctor of Philosophy; Engineering; Erythrocytes; FDA approved; Geometry; Goals; Hand; Harvest; Hematologic Neoplasms; Human; Immunophenotyping; Immunotherapy; improved; Injections; Lateral; Leukapheresis; leukemia; Leukocytes; Liquid substance; Malignant Neoplasms; Manufactured Materials; Methods; microchip; Microfluidic Microchips; Microfluidics; Molds; novel; operation; Output; Patients; performance tests; personalized medicine; Phase; Polymers; Population; Preparation; Procedures; Process; product development; Production; prototype; Reagent; Recovery; Refractory; Relapse; Running; Sampling; scale up; Silicon; Site; Small Business Technology Transfer Research; Solid; Stem cells; Sterility; Suspensions; T cell therapy; T-Lymphocyte; Technology; Testing; Therapeutic; Time; Translating; Work; young adult;

The goal of this Fast-Track STTR project is to develop a Deterministic Lateral Displacement (DLD) microfluidic device that can enrich white blood cells (WBCs) from a typical leukapheresis unit in 1 hr, for use in manufacturing cancer cellular immunotherapy. Chimeric antigen receptor T cell (CAR-T) therapy has been recommended for FDA approval to treat relapsed or refractory pediatric and young adult patients with B-cell acute lymphoblastic leukemia. There is a critical need for cost-effective automated methods to improve the efficiency and yield of large-scale enrichment of WBCs for use in manufacturing CAR-T and other cellular therapies. GPB is a pioneer in developing novel DLD microchips to process blood cells for cell analysis (19,26). GPB now proposes to develop, evaluate and commercialize a compact device in which an entire leukapheresis unit (up to 5x1010 WBCs in up to 300 ml) can be processed in a “Leuko-stack” of disposable single-use multi-channel DLD chips to produce in 1 hr a washed cell suspension that is enriched in WBCs and depleted of red blood cells (RBCs) and platelets (PLT). In Phase I, Aim 1 is to increase cell throughput through the current prototype chips by: 1) optimizing DLD chip design and operation to increase flow rate; 2) increasing throughput by stacking plastic chips and running them in parallel (“Leuko-stacks”); and 3) translating chip production to high-volume manufacturing material such as Cyclic Olefin Polymer (COP). Final Phase I milestones to proceed to Phase II are: 1) final chip design with a flow rate of at least 25 mL/hr via a single chip, at least 70% recovery of viable WBCs and immunophenotype- defined T-lymphocytes, and ability to process cells for 1 hr without clogging; 2) Leuko-stack of at least 6 chips run in parallel, with the same output as in #1; 3) combined increases in throughput via #1 and #2 sufficient to process a 300 ml leukapheresis unit in 1 hr; 4) confirmation that the chips can be produced from COP. In Phase II, Aim 2 is to build final prototype COP plastic chip-based microfluidic device capable of processing a leukapheresis sample at 300 mL/hr. Aim 3 is to test performance of prototypes from Aim 2 with leukapheresis aliquots and then full-size human leukapheresis samples. The final milestone of this project is to produce a set of commercial prototype Leuko-stacks that can process an entire 300-ml leukapheresis unit in 1 hr with at least 70% WBC and T-lymphocyte recovery, at least 90% depletion of RBCs, at least 80% depletion of PLTs, and at least 70% recovery of T-cell expansion capacity (as compared with the input samples) in significantly more than 50% of samples tested at 2 sites. The GPB Leuko-stack platform will preserve the advantages of DLD microfluidic cell processing over current methods, while massively increasing throughput rate and cell processing capacity, thus transitioning from analytic- to preparative-scale WBC enrichment for subsequent manufacture of CAR-T and other cell therapies.

Public Health Relevance Statement:
PROJECT NARRATIVE Targeted cellular immunotherapies for several hematologic malignancies, and potentially for solid cancers, are nearly at hand. In order to increase their availability, there is a critical need for better methods to efficiently and cost-effectively isolate large quantities of the white blood cells needed to manufacture the therapeutic cells in sufficient quantities. This project proposes to scale up a proprietary microfluidic cell processing technology to enrich white blood cells from leukapheresis harvests, the first step in engineering/manufacture of therapeutic chimeric antigen receptor T cells (CAR-T) that can cure leukemias and other cancers. Successful completion of this work will result in a well-characterized prototype for a white blood cell enrichment device that will be ready for product development and extensive testing and suitable to meet regulatory approval standards.

NIH Spending Category:
Bioengineering; Biotechnology; Cancer; Clinical Research; Gene Therapy; Genetics; Health Disparities; Hematology; Immunization; Immunotherapy; Minority Health; Vaccine Related

Project Terms:
Aliquot; anticancer research; Autologous; B-Cell Acute Lymphoblastic Leukemia; base; Blood; Blood Cells; Blood Platelets; Businesses; cancer cell; CAR T cell therapy; cell injury; Cell physiology; Cell Separation; Cell Therapy; cell type; Cells; Cellular immunotherapy; cellular targeting; Centrifugation; Childhood; chimeric antigen receptor T cells; Clinical; Clinical Trials; Collaborations; cost; cost effective; Cycloparaffins; design; Device or Instrument Development; Devices; Doctor of Philosophy; Engineering; Erythrocytes; FDA approved; Geometry; Goals; Hand; Harvest; Hematologic Neoplasms; Hematopoietic stem cells; Human; Immunophenotyping; Immunotherapy; improved; Injections; Lateral; Leukapheresis; leukemia; Leukocytes; Liquid substance; Malignant Neoplasms; Manufactured Materials; Methods; microchip; Microfluidic Microchips; Microfluidics; Molds; novel; operation; Output; Patients; performance tests; personalized medicine; Phase; Polymers; Population; Preparation; preservation; Procedures; Process; product development; Production; prototype; Reagent; Recovery; Refractory; Relapse; Running; Sampling; scale up; Silicon; Site; Small Business Technology Transfer Research; Solid; Sterility; Suspensions; T-Lymphocyte; Technology; Testing; Therapeutic; Time; Translating; Work; young adult