Chapter Four Simple high-throughput encoding of deep mutational scanning libraries by oligo-based golden gate assembly

ABSTRACT

Antibodies are versatile effector molecules of the adaptive immune system, distinguished by their capacity to recognise and engage diverse antigens with high specificity and affinity. The widespread adoption of antibodies, and their various formats, in diagnostics and therapeutics has been made possible through the co-operation of experimental and computational molecular engineering techniques. Functional screening of variant antibody libraries has proven essential for successful antibody development. Several protein display technologies have been deployed for this important screening phase, including phage display. Whilst one of the older approaches, phage display still offers many advantages over alternatives and synergises well with other advances in molecular biology techniques. This thesis demonstrates how phage display, when paired with modern techniques in novel ways, remains an extremely flexible and powerful tool for engineering and evaluating antibodies.