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The Use of Antigen-Coated Glass as a Specific Adsorbent for Antibody

G. Bonar Sutherland and Dan H. Campbell
J Immunol April 1, 1958, 80 (4) 294-298;
G. Bonar Sutherland
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Dan H. Campbell
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Discussion and Summary

Although the foregoing investigation demonstrated the applicability of antigen-coated glass surfaces for specific absorption much study must still be made on the quantitative aspects of the problem. The quantitative data obtained so far indicate that the method is not as efficient as might be expected. For example, in the ovalbumin antiovalbumin system using the tube method (see Table 1) 3.0 µg of ovalbumin absorbed only 18.0 µg of protein from the antiserum. Assuming this protein was antibody of 160,000 molecular weight, the mole ratio of antibody to antigen would be only 1.5. The relation of the amount of ovalbumin on the glass beads to the amount of antibody recovered from the columns indicated that this method with the present technique was less efficient than the tube method as might be expected.

Since small amounts of protein from normal sera were adsorbed to antigen surfaces, one might question the purity of antibody which was recovered. Furthermore, one would expect some coprecipitation of nonantibody protein from immune sera (e.g., complement). However, if one excepts the electrophoretic data, the purity of antibody from the glass bead columns must be at least of the order of 90–95% (including the nonprecipitating material).

However, the amount of antibody adsorbed is sufficient to give very significant analytical data and the method is applicable to the removal and detection of low concentration of antibody.

Footnotes

  • ↵This work was supported in part by the National Institutes of Health and in part by the Rockefeller Foundation.

  • Received November 13, 1957.
  • Copyright © 1958 by The American Association of Immunologists, Inc.

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The Journal of Immunology
Vol. 80, Issue 4
1 Apr 1958
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The Use of Antigen-Coated Glass as a Specific Adsorbent for Antibody
G. Bonar Sutherland, Dan H. Campbell
The Journal of Immunology April 1, 1958, 80 (4) 294-298;

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The Use of Antigen-Coated Glass as a Specific Adsorbent for Antibody
G. Bonar Sutherland, Dan H. Campbell
The Journal of Immunology April 1, 1958, 80 (4) 294-298;
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Print ISSN 0022-1767        Online ISSN 1550-6606