BME MS Defense: Meghan Jones

Development of a High-Throughput Screening Array for Immune Cells for the Advancement of HIV Research and Therapeutics

Supervised by Professor Lisa DeLouise

Abstract

Monoclonal antibody research is a multibillion-dollar per year market due to treatment successes and promises for cures of difficult pathogens, such as HIV. Sources of neutralizing antibodies are the antibody secreting cells (ASCs) of the immune system, or the B cells. In response to a pathogen, specific B cells become stimulated to produce the corresponding antibody to the pathogen. Microfabrication techniques are quickly becoming a sought after high-throughput screening tool to find single ASCs specific to a pathogen for improvement in the monoclonal antibody research field.

Microbubble (MB) array technology has been proven to be conducive to cell culture and microenvironment conditioning resulting in the accumulation of secreted factors. For these reasons, this thesis aims at exploiting MB technology for single cell culture and detection of ASCs. The seeding distribution within the MB array was characterized and modeled, allowing for future extrapolation of the seeding density or array size necessary to capture rare cell populations. Additionally, MB arrays display a relatively high total cell seeding necessary for enhanced detection of rare cells.

MB wells were also conducive to surface coatings for affinity capture of a cell of interest, which greatly reduces the background of nonspecific cells to retain single cell seeding of a specific cell. Once cell seeding characterization and enhanced capture efficiency were established, detection of a specific ASCs was performed. Unlike competitive technologies, MBs spherical well architecture allows for sustained cell culture over weeks leading to the detection of low antibody secreting cells. Detection of antibodies was validated by the identification of IgG secreting primary cells and tetanus specific ASCs within a high background of T cells. Coupled with high cell seeding and retention of the cell of interest, MB arrays show great promise as a high-throughput detection device.