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Enzyme Multiplied Immunoassay (EMIT)

Separation using the specificity of antibody - antigen binding and quantitation using enzyme reaction

Components of the EMIT Assay Method

Drug
Antibody
Substrate
Enzyme bound to drug

General Procedure for EMIT Assay

Mix sample containing drug with fixed quantity of enzyme bound drug, and antibody
Add substrate
Measure absorbance at 15 and 45 seconds after substrate addition
Quantitate by measuring enzyme-substrate reaction (by UV - visible spectroscopy)
Δ Absorbance from Reaction rate from Drug concentration
Non linear relationship between Δ Absorbance and Concentration
Determine standard curve

Fig 3.8.1 EMIT Assay Components in Action
Redrawn from: Pieper and Rutledge, Laboratory Techniques for Pharmacists, Upjohn 1989, page 40, figure 12

References

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