SBIR-STTR Award

The overarching technical objective of this proposal is to develop, prototype, and validate the HelioSAFETM sterility testing platform (Helio Sterility Assessment using Fluidics Enabled technology). To achieve this goal, we will pursue the following three Technical Objectives: Technical Objective 1. To develop and test the HelioSAFETM Cultivation Chip (HCC)TM, a novel microfluidic device that can rapidly concentrate microbial contaminants with minimum loss, incubate concentrated sample material in diverse culture media formulations, followed by rapid single-cell-resolution cell counting (before and after cultivation), to accurately and rapidly determine whether a sample contains viable microorganisms. Bacillus subtilis subsp. Spizizenii will be used as a model organism throughout this development process. Technical Objective 2. To expand HCC testing to other microorganisms and fully validate the performance of the HelioSAFETM system. We will expand the testing to Clostridium sporogenes, an anaerobe, to further validate the utility of HelioSAFETM. We will also finalize the performance characterization of the HCC and fix the design parameters towards future scale-up manufacturing. Technical Objective 3. To further develop the HelioSAFETM system as an in-line monitoring system. We will use sterile tubing to connect the HelioSAFETM chip to a simulated sterile biopharmaceutical production line and conduct periodic sampling and testing of the solution for microbial contaminants. This effort will demonstrate the feasibility of HelioSAFETM as an in-line sterility monitoring tool. The outcome of our pursuit of these objectives will be the development of a fully validated prototype HelioSAFETM sterility testing chip, determination of the performance of the HelioSAFETM system, demonstration of the feasibility of HelioSAFETM as an in-line fully automated sterility monitoring system, and identification of technical strengths and limitations (if any). In the future (e.g., Phase 2), we will optimize the HelioSAFETM HCC design with scale-up manufacturing in mind, demonstrate the feasibility of small-volume analysis on the chip, and develop the HCC chip reader (that includes a compact single-frequency impedance analyzer, compact OEM syringe pump, and pneumatic valve controller). In parallel, we will also test the following microbes from the DARPA BAA list as model microbial contaminants to further define the performance of the HelioSAFETM system: Candida albicans; Pseudomonas aeruginosa; Aspergillus brasiliensis; Staphylococcus aureus; and Bacillus subtilis subsp. spizizenii (spores).

There is a critical need within the Department of Defense (DoD) and the medical countermeasure (MCM) development community to rapidly manufacture biologics such as nucleic acid products and proteins (e.g., antigens, monoclonal antibodies) in varying quantities (gram to kg scales). These products can be used both as MCM and for other industrial processes such as enzyme-catalyzed reactions. Importantly, sterility testing is performed on MCMs to confirm that they are contaminant-free. Sterility testing is typically performed by taking a percentage of the total reagent or cellular inputs as well as the products to be tested in each manufactured batch. There are several limitations to current sterility testing methods, which are time-consuming and costly Therefore, a method that can provide rapid, small sample volume, fully automated, small footprint, low power requirements, and in-line sterility testing would constitute an ideal solution to overcome critical bottlenecks. Toward this end, HelioWave Technologies, LLC, proposes to advance the development of its HelioSAFETM system for rapid, label free automated sterility testing. This effort will involve validating the performance of the system using a panel of biological contaminants, including bacterial cells, bacterial spores, and fungi. In addition, the system will be tested using a variety of sample types derived from nucleic acid and protein therapeutic and medical countermeasure (MCM) workflows. In addition, the project will advance the development of the HelioSAFETM system toward FDA qualification and finalize the performance characterization of the device and detection hardware for scale-up manufacturing. Finally, Finally, we will subject the HelioSAFETM system to independent verification and validation testing to ensure it meets performance expectations. Taken together, these activities will deliver novel, rapid, and low-cost sterility testing that will positively impact biopharmaceutical manufacturing workflows.