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Title: Direct screening for antibody-antigen interactions
Author: Holt, L. J.
Awarding Body: University of Cambridge
Current Institution: University of Cambridge
Date of Award: 2001
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Array screening has been applied to the identification of antibodies from recombinant libraries, resulting in the identification of antibody-antigen interactions without the need for in vivo or in vitro antibody selection. In order to further increase the efficiency of array-based identification of molecular interactions, a novel two-dimensional arraying approach was invented and developed. Lines of biological sample from one repertoire were intersected with lines of sample from a second repertoire allowing interactions to be tested where the lines crossed. To test a given number of molecular pairs, the number of samples to be isolated, dispensed and stored and the time taken to create the array are greatly reduced when compared to one-dimensional arraying. The two-dimensional approach was applied to screening repertoires of antigens against repertoires of antibodies that bind to a variety of different proteins could be identified in a single experiment. In a second application of two-dimensional screening, large repertoires of antibodies were created on arrays by combining a few antibody heavy and light chains in all possible pairings. An array of 1.47 x 105 distinct antigen-binding sites was created by exhaustively combining 384 heavy chains and 384 light chains. It is likely that such repertoires, perhaps constructed from unselected antibody heavy and light chains, will provide a highly efficient method for the construction of antibody repertoires with which to analyse the human proteome.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available