A microarray format for multi-parameter blood group serology

Abstract

In this thesis multiple factors affecting probe-ligand interactions in a microarray format were investigated using blood group antigen and antibody sets for the development of a microarray based serology platform. Initial experiments focused on multiple parameters affecting immobilisation and functionality of monoclonal and polyclonal antibodies on various microarray surfaces. A range of critical parameters affecting protein-ligand interactions, such as efficient blocking of non-specific binding, detergent type, RBC concentration, reaction volume and mixing, were optimised in a series of experiments with labelled anti-species antibodies. Selected microarray surface and optimised spotting and reaction conditions were then used for studies of antibody-RBC antigen interactions in situ. A 'dual' solid-phase approach was investigated for blood group serology reactions where probe antibodies were immobilised on the microarray and target antigens were carried on the RBC surface, which can be considered as the second solid-phase. In summary this investigation has 1) established a microarray format and the reaction conditions for antibody-antigen interaction studies in blood group serology; 2) for the first time successfully exploited microarray format for comprehensive blood typing; 3) examined the novel technique of membrane fragment microarray immobilisation for a reverse, antibody screen reaction; 4) verified the findings and quantitatively characterised the interactions on Biacore, a surface plasmon resonance real-time detection system. This provides a strong basis for the development of microarray based multi-parameter blood group serology diagnostic platforms.EThOS - Electronic Theses Online ServiceGBUnited Kingdo