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An Enzyme‐Linked Antiglobulin Test to Detect Red Cell Globulins after Glutaraldehyde Fixation
Author(s) -
Greenwalt T.J.,
Dumaswala U.J.,
Siongco A.,
Domino M.M.
Publication year - 1992
Publication title -
vox sanguinis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.68
H-Index - 83
eISSN - 1423-0410
pISSN - 0042-9007
DOI - 10.1111/j.1423-0410.1992.tb01232.x
Subject(s) - glutaraldehyde , hemolysis , antibody , red cell , absorbance , chemistry , red blood cell , alkaline phosphatase , chromatography , globulin , enzyme , microbiology and biotechnology , biochemistry , immunology , medicine , biology
The purpose of this study was to develop an enzyme‐linked antiglobulin test (ELAT) for IgG on RBC without the hemolysis caused by the high pH of the alkaline phosphatase reaction. This was achieved by fixing the RBC with 0.05% glutaraldehyde after attachment of the antibodies. Assays using anti‐D reference standards demonstrated the sensitivity to be 1–2 ng of antibody as compared to 7.5 ng for the manual indirect antiglobulin test. The coefficients of variation of these assays ranged from 10.4 to 20.1%. The mean background absorbance at 405 nm of 105 normal RBC samples was 0.08+0.03 SD. There was an increase in sensitivity of the test after the fixed RBC were stored. Dilute glutaraldehyde stabilizes the RBC membrane and the antigen‐antibody linkage resulting in a more sensitive ELAT.