Antibody elution from red blood cells by chloroform and trichloroethylene

Abstract

Antibody elution by chloroform and trichloroethylene was compared with the Rubin ether method modified by Hughes-Jones for IgG antibodies and the Landsteiner and Miller heat method for IgM antibodies. After washing the cells, the chloroform and trichloroethylene method was done at 37 degrees C for 10 minutes. With this technique the eluate remained on the upper layer above the red blood cell stroma layer, while with Rubin's method the eluate remained below the cell stroma. The reported technique was as effective as the Landsteiner and Miller heat method for IgM antibodies, and a double volume of eluate was obtained.

Quantitative Evaluation of Elution Methods for Red Cell Antibodies

“…As far as the application of the ether method in general is concerned, the inferior results we have obtained could be due to the modifica tion of the original method [9] which omits the incubation of the ether-saline-red cell mixture for 30 min at 37 °C. This might partly explain the discrepancies of our find ings with those of prior publications [5,6], As is apparent from figure 2, eluates from in vitro sensitized RBC with alloantibodies were generally weaker than autoantibody eluates. It is known that stronger eluates can be produced if cells are exposed in vitro to larger amounts of serum.…”

“…Flowever, no single method produced the strongest eluate with all antibodies studied. We conclude that xylene and/or chloroform elution are valuable implementations in RBC serology, but, in selected instances, the classical elution methods will retain their importance.xylene [3,4] or chloroform [5,6], Since only semiquantitative comparisons of the elution efficiency of these two methods with the classical heat and ether elution have been reported in the literature [5], we were prompted to investigate these methods by means of a quantitative l25I-radioactive anti globulin test (125-I-RIAGT) for various types of RBC alio-and autoantibodies. …”

“…The heterogeneity of red cell antibodies has stimulated the search for an optimal elu tion technique [1][2][3][4][5][6]. Among the long list of recommended assays, heat elution has gained the widest acceptance both due to its simplicity and efficiency.…”

Primary anti‐D immunization by weak D type 2 RBCs

“…The tube was centrifuged for 10 minutes at 2500 × g , and the aqueous supernatant (eluate) was carefully harvested. The chloroform‐trichlorethylene technique 11 was also used.…”

Complement Is Not Activated in AB0-Haemolytic Disease of the Newborn: Two Exceptions

“…Immunoglobulins were eluted from cord blood red cells with chloroform and trichloro-ethylene [4], The specificity of the eluates was established in an indirect anti globulin test using adult A and B red blood cells. Maternal sera obtained soon after delivery were assayed for titre (indirect antiglobulin test) and subclasses of anti-A or anti-B IgG antibodies.…”