Development of single-cell sequencing based method for mAb identification and validation

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The overall goal of this project is to develop more cost-effective methods for generating monoclonal antibodies (mAb) against cancer biomarkers. While the traditional protocols for mAb generation used by companies such as MédiMabs have been established decades ago, recent technological developments have opened the door to new and potentially far more efficient methodologies. In this project, we will research the use of single cell DNA sequencing as a method to rapidly identify mAb producing cells without performing traditional hybridoma fusions. In order to do this, we will perform single cell RNA sequencing on B-cells from mice that have been immunized with an antigen of interest. When these cells are prepared for sequencing, the same antigen is added, however this version of the antigen also has a DNA barcode attached. When B cells that can recognize the antigen bind it, the attached DNA barcode is also added to the single cell sequencing reaction. This allows the simultaneous sequencing of both the barcode (if present) and the region encoding the antibody that can recognize the antigen. With this sequence information available, any antibodies of interest can be cloned into a new stable cell line for efficient production. At the same time, we will assess various approaches to optimize this part of the process, including rapid expression and translation of sequenced clones to allow their functional validation. These experiments will be done in the context of several biomarkers that have been identified in a subgroup of pediatric acute myeloid leukemia (AML) identified by the Wilhelm lab. Because of the numerous potential advantages of this approach with respect to time and cost of mAb production, there is both a strong scientific as well as economic value underpinning the proposed work.