Chapter 23
Pharmaceutical Analysis
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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 23.8.1 EMIT Assay Components in Action
Redrawn from: Pieper and Rutledge, Laboratory Techniques for Pharmacists, Upjohn 1989, page 40, figure 12
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