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Immunoassay of low to moderately abundant anionic proteins utilizing selective immobilization to chitosan-coated

Karen J Rees-Milton1, Tassos P Anastassiades

  • 1Division of Rheumatology and Arthritis Centre, Department of Medicine, Queen's University, Kingston, ON K7L3N6, Canada. reesmilton@sympatico.ca

Analytical Biochemistry
|September 29, 2009
PubMed

Insights

This study presents a novel two-stage immunoassay for quantifying low-abundance serum proteins. The method uses chitosan-coated plates for selective capture, achieving a sensitive detection limit for anionic serum proteins.

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Immunology

Background:

  • Quantifying low to moderately abundant serum proteins presents a significant challenge in biochemical analysis.
  • Immunoassays require exploiting specific antigen properties for effective capture and purification.

Purpose of the Study:

  • To develop and apply a two-stage immunoassay for the sensitive detection of anionic serum proteins.
  • To overcome limitations in current methods for serum protein quantitation.

Main Methods:

  • A two-stage immunoassay employing chromogenic and chemiluminescent substrates was designed.
  • Anionic serum proteins were selectively captured using positively charged chitosan-coated polystyrene plates at pH 6.1 with detergent.

Main Results:

  • The developed immunoassay demonstrated a detection limit of 0.1 microg/mL.
  • The assay exhibited a wide dynamic range of 1000-fold for protein quantitation.

Conclusions:

  • The described two-stage immunoassay effectively quantifies low to moderately abundant anionic serum proteins.
  • This method offers improved sensitivity and range for serum protein analysis in biochemical research.

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