Direct Comparisons between a Radio-Immune Antiglobulin Test, an Enzyme-Linked Antiglobulin Test, and a Haemagglutination Assay: Application to the Screening of Anti-RBC Sera and Monoclonal Antibodies

Abstract

Direct comparative studies between a radio-immune antiglobulin test (RIAT), an enzyme-linked antiglobulin test (ELAT) and a haemagglutination assay (HA) were carried out using anti-RBC hybridoma culture supernatants, monoclonal antibodies of established specificity, and immune mouse sera. The direct comparisons revealed that in many cases, RIAT and ELAT were slightly more sensitive than HA for the detection and study of anti-RBC antibodies. RIAT and ELAT detect non-agglutinating antibodies and sub-agglutinating concentrations of antibodies which would be missed if HA is used as the only screening test for hybridoma supernatants.

A review: the enzyme-linked antiglobulin test (ELAT) and its applications for reference laboratories

“…A solid-phase enzyme immunoassay was used by Francois-Gerard and Opret-Meunier sup (30) to subclass auto and alloantibodies. Bessos and Yul e sup (31) found the ELAT to be a good screening method for the detection of clones secreting antibodies, especially those that were not agglutinins. They also did not miss any monoclonal antibodies due to Automation of blood bank tests is an important area of investigation at present, and there are some tempting aspects of ELATs that could make them suitable for crossmatching and antibody screening.…”

Microimmunofluorescence Using Terasaki Plates and Direct Plate Freezing Method—Rapid and Reliable Screening System of Hybridomas

“…There are several methods for screening hybridomas; e.g., immunofluorescence staining (3), radioimmunoassay (RIA) (1), cytotoxic assay, enzyme-linked immunosorben t assay (ELISA) (2,5) and red cell adherence assay (11). The advantage of using immunofluorescence for hybridoma screening is that it reveals stable results and elucidates the specificities of antibodies against subcellular components, whereas a common binding assay such as RIA, can merely provide information about reactivity of the antibodies against whole cells.…”