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Park, Emily (2024) Isolation and characterisation of bovine ultralong antibodies to SARS-CoV-2 using phage display. PhD thesis, University of Nottingham.

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Abstract

The urgency for therapeutics in the arms race against SARS-CoV-2 increases with the emergence of each novel variant. Although current vaccines provide protection against severe disease in the majority of the population, there is still a great need for therapeutics for immunocompromised and severely ill patients. Broadly neutralising antibodies are highly adept at binding conserved epitopes, and, with a short developmental timescale, they are ideal candidates for antibody therapies.

In this study, consecutive immunisations were performed on two cows with Lineage A SARS-CoV-2 spike and peripheral blood mononuclear cells were isolated from the last bleed. Using this source, two bovine ultralong Fragment antigen binding (Fab) phage display libraries were constructed. Validation of the newly developed display libraries identified two Fab of interest, C13.2 and C13.7, that were subsequently cloned into a human antibody cassette and expressed in mammalian cells. Simultaneously, two rounds of biopanning were performed with the libraries against SARS-CoV-2 Lineage A trimer and SARS-CoV-2 BA.5 trimer, identifying several candidates which included C13.2 and C13.7. Full characterisation was performed on C13.2 and C13.7 through binding and neutralisation assays using spikes from different variants to assess breadth and potency. C13.2 and C13.7 were discovered to be broadly neutralising bovine antibodies. C13.2 potently neutralised pseudoviruses in vitro SARS-CoV-2 variants, C13.7 was less potent yet showed a greater breadth, neutralising pseudoviruses of SARS-CoV-2 variants and SARS-CoV-1.

C13.7 showed cross-reactivity to RBDs across three Sarbecovirus clades, warranting further investigation in vitro, in vivo and structural studies to determine the extent of binding and neutralisation, and ultimately the clinical potential, of this mAb. This approach identifies a pipeline for tapping into the bovine immune repertoire, which may be a powerful tool for combating rapidly emerging pathogens.

Item Type:	Thesis (University of Nottingham only) (PhD)
Supervisors:	Ball, Jonathan McClure, Patrick Tarr, Alexander Urbanowicz, Richard
Keywords:	SARS-CoV-2; Vaccines; Neutralising antibodies; Antibody therapies
Subjects:	Q Science > QR Microbiology > QR180 Immunology
Faculties/Schools:	UK Campuses > Faculty of Medicine and Health Sciences > School of Life Sciences
Item ID:	77643
Depositing User:	Park, Emily
Date Deposited:	16 Jul 2024 04:40
Last Modified:	18 Oct 2024 15:53
URI:	https://eprints.nottingham.ac.uk/id/eprint/77643

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