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Chip electrophoresis as a method for quantifying total microalbuminuria.

Owen T M Chan1, David A Herold

  • 1Department of Pathology, VA San Diego Healthcare System 92161, and University of California, San Diego School of Medicine, La Jolla, CA, USA. ochan@ucsd.edu

Clinical Chemistry
|September 23, 2006
PubMed
Summary

A new chip electrophoresis assay accurately measures total microalbuminuria, including nonimmunoreactive forms missed by traditional immunoassays. This method offers improved detection for diabetic nephropathy and cardiovascular disease risk assessment.

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Area of Science:

  • Clinical Chemistry
  • Analytical Biochemistry
  • Proteomics

Background:

  • Microalbuminuria is a key prognostic indicator for diabetic nephropathy and cardiovascular disease.
  • Traditional immunoassays may underestimate microalbuminuria by failing to detect nonimmunoreactive albumin forms.
  • Previous size-exclusion HPLC methods showed higher results due to interfering urine proteins.

Purpose of the Study:

  • To develop and validate a novel assay for quantifying total microalbuminuria, encompassing both immunoreactive and nonimmunoreactive forms.
  • To ensure the assay is free from interference by common urinary proteins.
  • To provide a more accurate measure of microalbuminuria for improved clinical prognostication.

Main Methods:

  • Utilized an automated chip electrophoresis system with microfluidic separation and fluorescent detection.

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  • Employed a chicken albumin internal calibrator for specimen normalization.
  • Analyzed albumin specimens without additional reducing agents.

    • Main Results:

    • The chip electrophoresis system demonstrated high linearity (R2=0.9997) and a low limit of detection (5 mg/L).
    • Interchip and intrachip coefficients of variation (CVs) were 3%-13%, with no significant impact from interfering agents.
    • The chip electrophoresis method consistently identified higher microalbuminuria concentrations than immunoassay and resolved albumin peaks from interfering proteins.

    Conclusions:

    • Chip electrophoresis accurately quantifies both immunoreactive and nonimmunoreactive albumin, unlike conventional immunoassays.
    • This method provides a simple, robust, sensitive, precise, and accurate approach for total microalbuminuria measurement.
    • The enhanced detection capabilities offer improved potential for assessing diabetic nephropathy and cardiovascular disease risk.