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Simultaneous determination of total IgE and allergen-specific IgE in serum by the MAST chemiluminescent assay system.

Authors
  • Cr, Brown
  • Kw, Higgins
  • Kelly A. Frazer
  • Lk, Schoelz
  • Jw, Dyminski
  • Va, Marinkovich
  • Sp, Miller
  • Jf, Burd
Type
Published Article
Journal
Clinical Chemistry
Publisher
American Association for Clinical Chemistry
Volume
31
Issue
9
Pages
1500–1505
Source
Frazer Lab
License
Unknown

Abstract

We have developed a chemiluminescent immunoenzymometric system. The first commercial application of this chemiluminescent assay (CLA) is the measurement of total IgE and allergen-specific IgE in human serum. The CLA system is a second-generation adaptation of the MAST RIA allergy profiling system. The MAST CLA system assay protocol consists of three steps: overnight incubation of serum, a 4-h incubation with enzyme-labeled antibody, and a 30-min chemiluminescent reaction, which produces a visible image (immunograph) on high-speed Polaroid instant film. The densities of the bands produced on the film are quantified with an inexpensive microprocessor-controlled infrared transmittance densitometer. The novel luminogenic substrates used yield a constant light output for over 2 h with an intensity at least 10-fold greater than that of commercial chemiluminescent reagents. The MAST CLA system exhibits sensitivity, specificity, and precision equal to that of the MAST RIA system (r = 0.96 for 40 serum samples analyzed with 25 allergens). As many as 35 different allergens per sample can be quantified in a single assay. The MAST CLA system requires no standard curve or volume-dependent pipetting steps, incorporates both positive and negative controls for each sample, and quantifies allergen-specific IgE at picomolar concentrations.

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