An Enzyme Linked Immunosorbent Assay (ELISA) for Detecting IgG Sensitized Erythrocytes

Abstract

This paper describes an Enzyme Linked Immunosorbent Assay (ELISA) for detecting IgG sensitized erythrocytes utilizing a commercially available anti-human IgG conjugated with alkaline phosphatase. Erythrocyte hemolysis in the assay was minimized by dissolving the p-nitrophenyl phosphate substrate in a carbonate-bicarbonate buffer. Nonspecific absorption of the enzyme conjugate to erythrocytes and glassware was reduced by adding 1% bovine serum albumin to wash solutions. Assay sensitivity was increased with greater concentrations of enzyme conjugate and erythrocytes in the incubation stage. The sensitivity of the described ELISA procedure is approximately equal to that of the standard antiglobulin test. Some possible future applications of ELISA in the blood bank are discussed.

Enzyme Immunoassay with Cell Suspensions: Studies on Reactions between Serum Antibodies and Cell-Surface Antigens

“…The results of our experiments with Rh anti serum and human RBC were in agreement with the previous reports [5,6] on the usefulness of EIAcs in this antibody-antigen system. We extended this obser vation by the finding that murine H-2 molecules, i.e., antigens of the major histocompatibility complex, could be detected on RBC.…”

“…Reactions o f Rh Antiserum with Human RBC Since EIAcs was originally described as a method for detection of alloantibodies to human RBC [5], in our first experiments the reactions of an Rh anti serum with humans RBC were tested. Simultaneously with EIAcs the standard Coombs' assay was per- The results of EIAcs with Rh antiserum and Rh-RBC were consistently negative.…”

A New Technique in Quantitative Immunohematology: Solid-Phase Kinetic Enzyme-Linked Immunosorbent Assay

“…ELISAs are widely used in biology and medicine, particularly with regard to the serology ofinfectious disease [10,11], Several applications to immunohematology have also been described [12][13][14][15][16], There are a number of advantages of ELISA testing rel ative to agglutination techniques. ELISA methods indirectly measure the primary in teraction of antibody with antigen.…”

Comparison between a Solid-Phase Low-Ionic-Strength Solution Antiglobulin Test and Conventional Low-Ionic-Strength Antiglobulin Test: Assessment for the Screening of Antierythrocyte Antibodies

“…However, the variable intensity of hemagglutination explains the subjectivity in interpreting the data and the difficulty in automating the process in contrast to other artificial agglutinations [8,10], Some publications have shown the feasibility of automating the antiglobulin test with Groupamatic and Immunoscan equipment [6,12], Solid-phase methods that used an enzyme-linked anti globulin test [3,16] or a solid-phase red cell adherence antiglobulin test [13,15] were described as an alternative to hemagglutination. Recently, Gordon and Ross [5] pub lished a solid-phase antiglobulin test in which the phase was made up of wet AB serum.…”