An avidin--biotin microELISA for rapid measurement of total and allergen-specific human IgE

J Immunol Methods. 1983 Feb 25;57(1-3):71-85. doi: 10.1016/0022-1759(83)90066-2.

Abstract

This paper describes an improved microtiter solid-phase enzyme immunoassay for the determination of total and allergen-specific human IgE. This assay technique is unique in its use of the avidin-biotin interaction to increase sensitivity. The avidin-biotin microtiter enzyme-linked immunosorbant assay (AB-microELISA) was performed in polyvinyl chloride microtiter plates using biotinylated anti-IgE and horseradish peroxidase (HRP)-avidin conjugate. This AB-microELISA technique enabled the quantitation of human serum IgE in the range of 0.1-5 ng/ml (10-500 pg/test) in less than 3 h. Total serum IgE, whether measured by the AB-microELISA or the paper radioimmunosorbant test (PRIST) was similar (correlation coefficient, r = 0.92). Further, the presence or absence of positive skin tests to 7 specific allergens determined in serum donors generally agreed with the presence or absence of allergen-specific IgE in their sera as measured by the AB-microELISA. The quantity of short ragweed allergen-specific IgE as determined by the AB-microELISA agreed with values obtained by the radioimmunosorbant test (RAST) (correlation coefficient, r = 0.89). No significant interference by ragweed-specific IgG (blocking antibody) was observed in the quantitation of allergen-specific IgE. The AB-microELISA is not only rapid and inexpensive, but also more sensitive than other published ELISA procedures and comparable to solid-phase radioimmunoassays in the quantitation of total and allergen-specific IgE.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allergens*
  • Antibody Specificity
  • Avidin
  • Biotin
  • Dose-Response Relationship, Immunologic
  • Enzyme-Linked Immunosorbent Assay
  • Horseradish Peroxidase
  • Humans
  • Immunoglobulin E / analysis*

Substances

  • Allergens
  • Avidin
  • Immunoglobulin E
  • Biotin
  • Horseradish Peroxidase