Antibody production using genetic immunization

Extract: Antibodies are key tools in proteomics. Their high specificity and affinity allow them to be used to measure individual proteins in complex mixtures, and most importantly to provide quantitative information. There is now great interest in generating proteome-wide sets of antibodies in order to discover biomarkers of disease and to enhance drug development. Antibodies are traditionally produced by injecting a pure form of the protein mixed with an adjuvant into an animal. The pure protein is usually produced either by overexpression in a host such as E. coli and purifying it using a fused tag (e.g., GST, 6-histidines), or by synthesizing small peptide sequences from the whole protein. The breakthrough effort in the last decade of determining the sequences of entire genomes has discovered tens of thousands of new proteins. Antibodies are now the major limiting factor in quantitative proteomics. The protein-based technology has proven too slow and expensive to make it feasible to produce antibodies on the scale now needed by the pharmaceutical and biotechnology industries.