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Detection of Cell-Bound Immunoglobulins by a Radioisotopic Micro-Mixed Hemadsorption Reaction with Technetium-99m-Labeled Erythrocytes

Gary W. Wood and Rolf F. Barth
J Immunol March 1, 1975, 114 (3) 944-949;
Gary W. Wood
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Abstract

The mixed hemadsorption (MHA) reaction detects antibodies reactive with cell surface antigens by means of antiglobulin-coated indicator erythrocytes. We have developed a radioisotopic modification which employs sheep erythrocytes (SRBC) that have been prelabeled with technetium-99m (99mTc), a high specific activity metastable γ-emitter of short half life. The 99mTc MHA reaction was performed on human and murine cells cultured in Microtest II plates with six replicate wells per serum dilution. Antibody activity in species-specific xenoantisera and mono- and polyspecific alloantisera was detected in high titer. The sensitivity of 99mTc micro-mixed hemadsorption was 2 times that of the visual assessment of mixed hemadsorption, 100 to 200 times that of the 125I-mixed antiglobulin reaction and 500 to 1000 times more sensitive than indirect immunofluorescence. The assay system was applied successfully to confirm the species of origin of a panel of previously karyotyped human and mouse cell lines. Our results indicate that the 99mTc micro-mixed hemadsorption method is a rapid, sensitive, quantitative test for the detection of cell surface antigens and membrane reactive antibodies.

Footnotes

  • ↵3 Postdoctoral Fellow, United States Public Health Service, 1972–1973.

  • ↵2 This investigation was supported by Public Health Service General Research Grant 5 SO4 RRO6147, Grants AI 09947 and CA 13190 from the National Institutes of Health, and DRG 1075 B, Damon Runyon Memorial Fund.

  • Received September 10, 1974.
  • Copyright © 1975 by The American Association of Immunologists, Inc.

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The Journal of Immunology
Vol. 114, Issue 3
1 Mar 1975
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Detection of Cell-Bound Immunoglobulins by a Radioisotopic Micro-Mixed Hemadsorption Reaction with Technetium-99m-Labeled Erythrocytes
Gary W. Wood, Rolf F. Barth
The Journal of Immunology March 1, 1975, 114 (3) 944-949;

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Detection of Cell-Bound Immunoglobulins by a Radioisotopic Micro-Mixed Hemadsorption Reaction with Technetium-99m-Labeled Erythrocytes
Gary W. Wood, Rolf F. Barth
The Journal of Immunology March 1, 1975, 114 (3) 944-949;
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