Variability of IgE protein measurement in cell-culture supernatants: Results from a multicenter collaborative study

Ricki M. Helm, Rebecca H. Buckley, N Franklin Adkinson, Diane L. Squillace, Gerald J. Gleich, John W. Yunginger

Research output: Contribution to journalArticle

Abstract

The sensitivity, specificity, and precision of immunoassays for quantitation of IgE in cell-culture supernatants were tested in a multicenter trial involving 22 laboratories. Fourteen coded test samples included cell-culture medium alone, culture medium with varying concentrations of polyclonal or myeloma IgE, and medium from unstimulated or pokeweed mitogen (PWM)-stimulated peripheral blood mononuclear cell (PBMC)-culture supernatants. Two laboratories reported measurable IgE in non-IgE-containing control samples. Although the IgE content of a single 0.50 ng%ml polyclonal IgE sample should have been measured easily by the claims of all laboratories, only 13 laboratories measured IgE in this sample in 22 of the 48 assays. Most laboratories could measure both polyclonal and myeloma IgE at 5.0 nglml; however, the IgE determinations for the myeloma proteins were nearer the predicted value. Although 15 laboratories found measurable IgE in the PWM-stimulated PBMC-culture supernatants from a single nonatopic donor, the levels did not differ significantly from that measured in unstimulated PBMC-culture supernatants. Three laboratories reported considerably higher IgE levels in the PWM-stimulated PBMC-culture supernatants than in other PBMC-culture supernatants. Only 13 laboratories could quantitate IgE in each of coded duplicate samples containing 0.50 nglml polyclonal IgE. These findings indicate a wide variability in the sensitivity, specificity, and precision of assays used to quantitate low levels of IgE protein. Investigators should be encouraged to make greater use of existing national and international reference materials in the standardization and performance of their IgE immunoassays.

Original languageEnglish (US)
Pages (from-to)880-890
Number of pages11
JournalThe Journal of Allergy and Clinical Immunology
Volume77
Issue number6
DOIs
StatePublished - 1986

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Immunoglobulin E
Multicenter Studies
Cell Culture Techniques
Proteins
Blood Cells
Pokeweed Mitogens
Immunoassay
Culture Media
Myeloma Proteins
Sensitivity and Specificity
Research Personnel

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

Cite this

Variability of IgE protein measurement in cell-culture supernatants : Results from a multicenter collaborative study. / Helm, Ricki M.; Buckley, Rebecca H.; Adkinson, N Franklin; Squillace, Diane L.; Gleich, Gerald J.; Yunginger, John W.

In: The Journal of Allergy and Clinical Immunology, Vol. 77, No. 6, 1986, p. 880-890.

Research output: Contribution to journalArticle

Helm, Ricki M. ; Buckley, Rebecca H. ; Adkinson, N Franklin ; Squillace, Diane L. ; Gleich, Gerald J. ; Yunginger, John W. / Variability of IgE protein measurement in cell-culture supernatants : Results from a multicenter collaborative study. In: The Journal of Allergy and Clinical Immunology. 1986 ; Vol. 77, No. 6. pp. 880-890.
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