Antibodies in proteomics II: screening, high-throughput characterization and downstream applications

• Published in: Trends in biotechnology (Regular ed.) 2003-07, Vol.21 (7), p.312-317
• Main Authors: Bradbury, Andrew, Velappan, Nileena, Verzillo, Vittorio, Ovecka, Milan, Chasteen, Leslie, Sblattero, Daniele, Marzari, Roberto, Lou, Jianlong, Siegel, Robert, Pavlik, Peter
• Format: Article
• Language: English
• Subjects: Antibodies - immunology; Antibodies - isolation & purification; Antibodies - metabolism; Automation; Binders; Biological and medical sciences; Biotechnology; Cloning; Drug Evaluation, Preclinical - methods; Fundamental and applied biological sciences. Psychology; Genes; Genetic engineering; Immobilization of enzymes and other molecules; Immobilization techniques; Immunization; Immunoassay - methods; Libraries; Ligands; Methods; Methods. Procedures. Technologies; Peptide Library; Peptides; Protein Array Analysis - methods; Protein Engineering - methods; Proteins; Proteomics; Proteomics - methods
• ISSN: 0167-7799; 1879-3096
• DOI: 10.1016/S0167-7799(03)00117-3

There are many ways in which the use of antibodies and antibody selection can be improved and developed for high-throughput characterization. Standard protocols, such as immunoprecipitation, western blotting and immunofluorescence, can be used with antibody fragments generated by display technologies. Together with novel approaches, such as antibody chips and intracellular immunization, these methods will yield useful proteomic data following adaptation of the protocols for increased reliability and robustness. To date, most work has focused on the use of standard, well-characterized commercial antibodies. Such protocols need to be adapted for broader use, for example, with antibody fragments or other binders generated by display technologies, because it is unlikely that traditional approaches will provide the required throughput.