Immunoassays: Biological Tools for High Throughput Screening and Characterisation of Combinatorial Libraries

Abstract

In the demanding field of proteomics, there is an urgent need for affinity-catcher 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 characterisation of specific antibodies represent a key issue to meet success. Alternatively, it is expected that new, well-characterised affinity reagents generated in rapid and 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 (HTS) technologies have become essential for the generation and identification of robust affinity reagents from biological combinatorial libraries and the lead 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 ligand-receptor 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 low-molecular-weight bio-inspired mimic compounds. No matter what the ligand source, selection and characterisation of leads is a most relevant task. Immunological assays, in microtiter plates, biosensors or microarrays, 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.

Cavity-Enhanced Immunoassay Measurements in Microtiter Plates Using BBCEAS

“…The results obtained in the studied concentration range were significant as they showed the ability of the system to measure higher analyte concentrations and thus broaden its applicability. [1][2][3][4][5] in the concentration range 0 ng mL -1 to 61.6 ng mL -1 . The average CEF and the standard deviation of the measurement is also listed.…”

“…They are based on the detection of highly specific antibody antigen reactions with key characteristics of, accuracy, precision, limit of detection, measurement range, and cost. With the availability of a wide range of disease biomarkers and increased use of expensive reagents an ELISA can be costly to use if several biomarkers need to be measured in an investigation [1] . The most commonly used ELISA detection method involves colorimetric detection through the use of visible wavelength absorption spectroscopy with an appropriate microplate reader.…”

“…With the availability of a wide range of disease biomarkers and increased use of expensive reagents, an ELISA can be costly to use if several biomarkers need to be measured in an investigation. 1 The most commonly used ELISA detection method involves colorimetric detection through the use of visible wavelength absorption spectroscopy with an appropriate microplate reader. There are, however, a variety of other formats including simple dip strip ELISA.…”

A Surface Plasmon Resonance Sensor for the Detection of Deoxynivalenol Using a Molecularly Imprinted Polymer

“…Recently, immunosensors that use the antigen-antibody reaction have also been used for the rapid and easy determination of DON [16,17]. Nevertheless, these methods are expensive and affected by physical and chemical conditions.…”

Building Addressable Libraries: Site-Selective Suzuki Reactions on Microelectrode Arrays

“…Addressable microelectrode arrays , are intriguing tools for probing interactions between small molecules and biological receptors , because they enable monitoring of the interactions in “real-time” . For this reason, we have been developing synthetic methods for site-selectively building molecules proximal to the electrodes in a microelectrode array. , The long-term goal of the work is to use the arrays as a platform for small molecular libraries containing engineered, semirigid peptidomimetics that are useful for probing the conformational requirements of protein receptors.…”