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Related Experiment Videos

Logit-log calibration curves for EMIT assays.

D N Dietzler, M P Leckie, C R Hoelting

    Clinica Chimica Acta; International Journal of Clinical Chemistry
    |January 24, 1983
    PubMed
    Summary

    Accurate enzyme-multiplied immunoassay (EMIT) data analysis requires determining delta A infinity. An antibody-free substrate reagent reliably determined this value, overcoming limitations of other methods.

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    Area of Science:

    • Biochemistry
    • Analytical Chemistry
    • Clinical Chemistry

    Background:

    • Enzyme-multiplied immunoassay (EMIT) data analysis often uses logit-log or four-parameter logistic models.
    • Accurate application of these models necessitates determining the change in absorbance at infinite drug concentration (delta A infinity).
    • Variability in delta A infinity with equipment and reaction conditions complicates its determination and limits model applicability.

    Purpose of the Study:

    • To evaluate two simplified methods for accurately determining delta A infinity in EMIT assays.
    • To identify a reliable method for delta A infinity determination applicable across different EMIT assays and conditions.

    Main Methods:

    • Assessed delta A infinity determination using a cross-kit reaction method with reagents from different EMIT kits.

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  • Evaluated delta A infinity determination using an antibody-free substrate reagent based on direct analysis.
  • Tested methods with EMIT assays for phenobarbital, primidone, phenytoin, carbamazepine, ethosuximide, and theophylline.

    • Main Results:

    • The cross-kit reaction method sometimes yielded erroneously low delta A infinity values due to unexpected cross-reactions.
    • The antibody-free substrate reagent consistently enabled accurate determination of delta A infinity.
    • The antibody-free substrate reagent proved more reliable than the cross-kit method for various EMIT assays.

    Conclusions:

    • The antibody-free substrate reagent method is a robust and accurate approach for determining delta A infinity in EMIT assays.
    • This method overcomes the limitations associated with equipment and reaction condition variability.
    • Accurate delta A infinity determination facilitates broader and more reliable application of logistic models in EMIT data analysis.