Immunological Assays: Biotools for High Throughput Screening and Characterisation of Combinatorial Libraries
Pp. 130-158 (29)
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.
Antibody engineering, combinatorial chemistry, biological libraries,
chemical synthesis, high throughput screening, immunoassays, ELISA.
BB – Institute for Biotechnology and Bioengineering, Centro de Engenharia Biológica e Química, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001 Lisboa, Portugal.