ABSTRACT
In vitro diagnostic (IVD) platforms provide rapid and accurate determination of disease status. The clinical performance of antibody-based diagnostic platforms is paramount as the information provided often informs the medical intervention taken and, ultimately, the patient's outcome. Breaking down such an immuno-IVD device into its component elements, the biorecognition entity is key to the analytical specificity of the test. Furthermore, tailored optimisation of the antibody is often necessary to impart the desired biophysical properties for the specific application. This tailoring is now widely facilitated by advances in combinatorial approaches to antibody generation, molecular evolution strategies and the availability of truly high-throughput (HT), refined surface plasmon resonance-based screening tools. In this paper, we demonstrate a rational, knowledge-driven approach to the generation of epitope-specific antibodies for the early detection of cardiovascular disease, discuss the merits of the approaches taken and offer a perspective on HT strategies to mining large antibody libraries. These results highlight the expedience of such methodologies for the development of truly superior cardiovascular disease biorecognition elements.
Subject(s)
High-Throughput Screening Assays/methods , Immunologic Tests/methods , Peptide Library , Recombinant Proteins/metabolism , Single-Chain Antibodies/metabolism , Troponin I/analysis , Adjuvants, Immunologic , Amino Acid Sequence , Animals , Biomarkers/analysis , Biomarkers/metabolism , Chickens , Female , Hemocyanins , Humans , Hybridomas , Mice , Mice, Inbred BALB C , Molecular Sequence Data , Mutation , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Single-Chain Antibodies/genetics , Single-Chain Antibodies/immunology , Surface Plasmon Resonance , Troponin I/immunology , Troponin I/metabolismABSTRACT
A simple cell suspension enzyme-linked immunosorbent assay system, which can be used to test the reactivity of antibodies with cells, is described. Such a system is of particular use with cells that are difficult to fix to microtitre plates or when measuring antigens which are labile to fixation techniques.
