Background: Gentamicin residue in the production of biological samples may lead to severe allergic reactions in certain groups. Thus it is strictly controlled in many countries in the world and the analyte needs to be measured in biological preparations before release. This ELISA can be used in the production of therapeutic drugs and vaccines, to accurately quantify drug residues and ensure quality and purity of the final product. Current method of drug residue testing such as HPLC or GC-MS are with sample prep not only complex and expensive, but also difficult and time consuming. ELISA speed up and simplify the analysis without losing accuracy.
Principle: The method employs sandwich ELISA technique. Monoclonal antibodies to Gentamicin are pre-coated onto microwells. Samples and standards are pipetted into microwells and Gentamicin present in the sample are bound by the antibodies competing with Gentamicin labeled HRP. Post incubation a complex is formed. After washing the microwells in order to remove any non-specific binding, the substrate solution (TMB) is added to microwells and color develops proportionally to the amount of Gentamicin in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.
