Why does ELISA involve various degree of non-specific binding?

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Why does ELISA involve various degree of non-specific binding?

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Binding to the receptor of interest is called specific binding, while binding to the other sites is called nonspecific binding. In most cases, the bulk of nonspecific binding represents some sort of interaction of the ligand with membranes. The molecular details are unclear, but nonspecific binding depends on the charge and hydrophobicity of a ligand – but not its exact structure. Nonspecific binding is usually (but not necessarily) proportional to the concentration of radioligand (within the range it is used). Add twice as much radioligand, and you’ll see twice as much nonspecific binding. Nonspecific binding is detected by measuring radioligand binding in the presence of a saturating concentration of an unlabeled drug that binds to the receptors. Under those conditions, virtually all the receptors are occupied by the unlabeled drug so the radioligand can only bind to nonspecific sites. Subtract the nonspecific binding at a particular concentration of radioligand from the total bindin

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