Quantitation of red cell–bound IgG, IgA, and IgM in patients with autoimmune hemolytic anemia and blood donors by enzymelinked immunosorbent assay

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Immunohematology

American National Red Cross

Subject: Medical Laboratory Technology

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ISSN: 0894-203X
eISSN: 1930-3955

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VOLUME 19 , ISSUE 2 (June 2003) > List of articles

Quantitation of red cell–bound IgG, IgA, and IgM in patients with autoimmune hemolytic anemia and blood donors by enzymelinked immunosorbent assay

Antonio A. Bencomo / Martha Díaz / Yalile Alfonso / Odalys Valdés / María E. Alfonso

Keywords : enzyme-linked immunosorbent assay, autoimmune hemolytic anemia, IgG, IgA, and IgM immunoglobulins, RBCs, quantitative method

Citation Information : Immunohematology. Volume 19, Issue 2, Pages 47-53, DOI: https://doi.org/10.21307/immunohematology-2019-474

License : (Transfer of Copyright)

Published Online: 14-October-2020

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ABSTRACT

This paper describes an enzyme immunoassay for the quantitative determination of IgG, IgA, and IgM immunoglobulins on RBCs. Ether eluates made from RBCs were followed by an enzyme-linked immunosorbent assay of immunoglobulin concentration. Calibration curves were derived from immunoglobulin standards and the number of molecules of each isotype per RBC was calculated. The assay was carried out in 200 healthy blood donors and 62 patients with warm autoimmune hemolytic anemia (AIHA), two of them with a negative DAT. For healthy blood donors, mean values were 58 IgG, 16 IgA, and 3 IgM molecules per RBC. For patients with a positive DAT, the mean values were 3435 IgG, 157 IgA, and 69 IgM molecules per RBC. An increased level of IgA was found in 12 patients without IgA autoantibodies demonstrable in RBC eluates. Increased IgG levels were also observed in patients with a negative DAT and, in one case, an increased level of IgA was also found. The enzyme-linked immunosorbent assay using ether eluates is a sensitive method for quantitating RBC autoantibodies in patients with AIHA as well as immunoglobulins bound to RBCs in healthy individuals.

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