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From the Laboratory of Chemical Pharmacology, National Cancer Institute,,2 Bethesda, Maryland
Abstract
A rapid and reproducible assay method for cytolytic antibodies is described. The procedure is founded on the observation that trypsin digests cells after their exposure to antibody and complement.
In practice cells are exposed to various dilutions of antiserum plus complement and injured cells digested by trypsin and the surviving cells are counted. For rapid and accurate counting of cells, the Coulter electronic particle counter was utilized. The electronic impulses registered by the products of the tryptic digestion were negligible compared to those registered by intact cells.
In a model system, a horse antiserum against mouse Sarcoma 37 ascites cells was assayed. The cytolytic titer of the serum was expressed as the dilution resulting in lysis of 50% of cells. Assay of antiserum by this method was in close agreement with that obtained by the dye exclusion test.
In the range from 24 to 78% lysis, the standard deviation was less than ±3% lysis. Even the low levels of natural antibodies against sarcoma cells present in normal human serum were found to be well within the range of measurement of this method.
Footnotes
1 Present address: Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland.
2 National Institutes of Health, USPHS, United States Department of Health, Education, and Welfare.
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