Chapter 5

Immunological Assays: Biotools for High Throughput Screening and Characterisation of Combinatorial Libraries

Maria Â. Taipa


In the demanding field of proteomics, there is a great need for affinitycatcher molecules to implement effective and high throughput methods for analysing the human proteome or parts of it. Antibodies have an essential role in this endeavour, and selection, isolation and characterization of specific antibodies represent a key issue to meet success. Alternatively, it is expected that novel affinity reagents generated in fast, cost-effective manners will also be used to facilitate the deciphering of the function, location and interactions of the high number of encoded protein products. Combinatorial approaches combined with high throughput screening technologies have become essential for the generation and identification of robust affinity reagents from biological combinatorial libraries and the discovery of active/mimic molecules in large chemical libraries. Phage and yeast display provide the means for engineering a multitude of antibody-like molecules against any desired antigen. The construction of peptide libraries is commonly used for the identification and characterisation of ligandreceptor specific interactions, and the search for novel ligands for protein purification. Further improvement of chemical and biological resistance of affinity ligands encouraged the “intelligent” design and synthesis of chemical libraries of lowmolecular- weight bio-inspired mimic compounds. No matter what the ligand source, selection and characterization of leads is a most relevant task. Immunoassays are a biological tool of inestimable value for the iterative screening of combinatorial ligand libraries for tailored specificities, and improved affinities. Particularly, enzyme-linked immunosorbent assays are frequently the method of choice in a large number of screening strategies, for both biological and chemical libraries.

Total Pages: 130-158 (29)

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