UBC Theses and Dissertations
A microfluidic approach for antibody screening from single cells with numerical methods for modelling solid-substrate capture Lewis, Daniel Jacob
Antibodies are the fastest growing class of therapeutics and are also ubiquitous reagents in diagnostic and research applications. Despite the advance of many applications of antibodies in basic and applied research, the majority of novel antibody discovery and screening still typically relies on the costly, time-consuming and ine cient hybridoma approach; the development of improved approaches for the high-throughput screening and selection of monoclonal antibodies is thus of high interest. In this thesis a novel approach combining the concentration enhancement and sample manipulation bene ts of polydimethylsiloxane (PDMS) multilayer micro uidics devices is validated for the high-throughput prescreening of large samples of primary antibody secreting cells. Dual functionalized beads are co-incubated with antibody secreting cells to capture both antibodies and their associated messenger RNA (mRNA) transcripts to couple sequence with a nity information in downstream uorescence activated cell sorting (FACS). This is done in a parallel way for simultaneous cell processing over a large planar array. The device is validated with quantitative reverse transcription polymerase chain reaction (RT-qPCR) to verify successful complementary DNA (cDNA) synthesis on the bead surface and that the beads retain functional antibodies throughout the incubation and reverse transcription process. As part of this work, I also present a numerical modelling pipeline to design and characterize performance of di usion based solid substrate cell secretion assays in micro uidic chambers prior to time-consuming fabrication and experimentation.
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