Diagnostic Challenges: Advancing Assay Design

Abstract

Prologue: A unique aspect of the Professional Science Masters Applied Bioscience program is to immerse future graduates into the fields of scientific research inclusive of both business and basic science. At the culmination of the program, an 8 credit hour internship requirement proved to be an invaluable opportunity for me to experience a broad range of scientific inquiry. Throughout the duration of the internship, I was able to learn and master laboratory techniques, adhere to federal and state regulations, practice communicating my results, and ultimately be a contributing member of a collaborative team. In return, I aided my supervisors by completing projects and assigned tasks that was of great value and/or interest to the company. Internships can be done at one company or split to be completed at two separate industries. I was fortunate enough to be able to experience two companies, one focusing more on research and assay development (MSDx) and the other focusing on diagnostic tools to add to their testing services (Pharos Dx). The first internship was with MSDx, Inc., a research company aiming to provide diagnostic solutions for neurodegenerative disorders. The researchers were interested in moving a first generation assay to a second generation assay for greater specificity in detecting tau, a protein which is primarily located within the brain and a proven marker for brain degeneration [10, 11]. Their first generation assay, an indirect ELISA, is compatible with their current anti-tau monoclonal and polyclonal antibodies. My responsibilities were to search for additional anti-tau antibodies that could be used as pairs in a sandwich assay and would detect an epitope within the tau amino acid sequence of interest (c-terminus of the protein). At the conclusion of the internship, I created a table of commercially available antibodies and provided it to the company (table 1 within the paper). I was also able to demonstrate how the first generation assay worked to model the first steps that would be taken when testing a new antibody for assay development (figure 4). The second internship took place at Pharos Dx., which is a diagnostic company utilizing liquid-chromatography mass-spectrometry as part of their testing services. Pharos Dx was looking to offer microsampling devices to their clientele as it would help veterinarians to easily collect samples from animals. Microsampling has a disadvantage with the hematocrit where an unknown volume of blood would affect the overall spreading and homogeneity of the sample across the filter paper [43, 46]. My responsibility during this internship was to research the various microsampling tools currently out on the market. With the amount of published papers available, my objective was to sort through the validated and completed experiments to make a recommendation to Pharos Dx regarding the microsampling device to pursue and explore further. At the conclusion of the internship, I summarized my findings (presented within this paper) and suggested Pharos Dx begin confirmatory testing of using a potassium based algorithm to remove the hematocrit bias associated with dried blood spot cards [52, 64]. In conclusion, each internship was a full experience of new content and laboratory techniques. I gained a greater appreciation for what scientists have to go through to create commercial products to be used in disease detection and monitor overall health. With MSDx, Inc., I was able to learn a substantial amount about brain disorders and the recent findings (2015) of lymphatic vessels in the brain [5, 6]. With Pharos Dx., even though the focus was researching microsampling for a diagnostic tool, I learned more about the regulatory side of research and how to trust published results based on the validation of the procedures that were run. It was my goal that I would be able to contribute tangible products for these companies, as both companies truly helped me grow as a scientist.