BME MS Thesis Defense Seminar: Bryan Bobo
Friday, April 11, 2014
MC 2-6408 (K-207)
Development of Microbubble Array for High Throughput Screening of Cells by Their Secreted Factors
Advisor: Lisa DeLouise
Monoclonal antibody (mAb) production is a multibillion dollar global industry. The therapeutic potential of mAbs makes them an ideal tool in both clinical and research applications due to their ability to recognize and bind specific epitopes. While mAbs are a silver bullet in the medical industry their utility is overshadowed by the cost associated with their production which relies on the ability to identify small minorities of mAb producing cells called hybridomas out of a heterogeneous population. To address concerns with suboptimal screening methods for cell sorting many groups have begun to investigate the use of microarrays for the purpose of mAb production and detection of antibody secreting cells (ASCs).
To surmount problems associated with mAb production Microbubble (MB) arrays were developed and proof-of-concept assays were outlined to confirm the ability of MBs to sort and detect ASCs within a heterogeneous population. This thesis herein presents methods to enhance the fluorescent signal associated with ASCs. The ability to enhance singles from ASCs enabled the rapid screening of MB arrays for antigen specific signal detection as well as the eventual IgG detection of primary B cells from human donors. Using the methods mentioned herein, rapid image analysis protocols were developed for the increased image processing of large MB arrays (>2500 MBs). These techniques have facilitated the screening of larger cell cohorts faster and more efficiently than ever before, enabling the detection and recovery of cells present at a 1:50 dilution.
The results presented within this thesis further substantiate the utility of the MB arrays for use in rapid cell screening applications such as hybrioma/mAb production. Due to their simplicity and low manufacturing cost MB arrays show great promise in various cell sorting applications.