Elution of HL‐A‐Specific Antibodies from Platelets1

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Abstract
The preconditions and kinetics of absorption and elution of HL‐A specific antibodies from platelets, their antibody binding capacity (‘activity’) and the stability of HL‐A receptors on platelets were studied. It was found that the acid elution technique is the only useful method as compared to the ether or heat elution. The optimal ratio of absorbed serum to eluate volume, platelet number, incubation temperature and time were evaluated. At pH 3.0–3.5 (range tested between pH 1.5 and 5.0) the strongest eluate activity was obtained. The fixation of HL‐A specific antibodies to platelet receptors was not reduced by repeated washings with buffered saline, hypo‐ and hyperosmolaric salt solutions (range 0.077 to 2.464 mol NaCl). The capacity of antigenic receptors of platelets to bind complement‐fixing HL‐A antibodies remained unaltered after treatment with distilled water, hypo‐ and hyperosmolaric salt solutions (range as above) and incubation between pH 5 and 9. Freezing (−20°C), heating (56°C) and short incubation at pH 10 moderately altered HL‐A receptors on platelets, which clearly deteriorated at pH 3 or after incubation at 100°C. A standardized elution procedure is recommended.