Abstract
Proteomics in Diagnosis: Past, Present and Future
Highlights
METHOD DESCRIPTION■ Separation of complex proteins via 2D gel electrophoresis based charge and size ■ Major protein identification by Mass spectrometry (MS) ■ Detects about 2000-2500 spots/gel
Proteome was coined by Marc Wilkins, which he used for the entire complement of protein expressed by a genome, cell, tissue or organism
Cellulose acetate/citrate agarose gel electrophoresis involves separating negatively charged proteins on an alkaline pH (8.2-8.6), due to variability in amino acid composition the charge of two molecules varies and two proteins with similar weight can be resolved at different locations, this needed to be confirmed with running the gel in acidic conditions in which samples are not being resolved on alkaline gel
Summary
■ Separation of complex proteins via 2D gel electrophoresis based charge and size ■ Major protein identification by MS ■ Detects about 2000-2500 spots/gel. Cellulose acetate/citrate agarose gel electrophoresis involves separating negatively charged proteins on an alkaline pH (8.2-8.6), due to variability in amino acid composition the charge of two molecules varies and two proteins with similar weight can be resolved at different locations, this needed to be confirmed with running the gel in acidic conditions (pH 6.0- 6.2) in which samples are not being resolved on alkaline gel In this method citrate was added to agar to make the surface acidic and the charge of amino acids varied according to their respective isoelectric points. Immunological techniques especially those based on selected antibodies for the detection of plethora of blood proteins identified as biomarkers are the most dominant diagnostic tools in use at present. Among infectious diseases several viral diseases such as HIV infection, Lyme diseases and recently emerging Ebola virus are diagnosed with the help of ELISA
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