First Results in the prevaluation of the human whole blood assay for pyrogens in biological pharmaceuticals

When establishing the whole blood assay for pyrogens in our laboratory Me d id several investigations on the standard pyrogen to be used Because of the hydrophobicity, of the lipopolysaccharid (LPS) it is difficult to get a ho mogeneous solution. Therefore we used ultrasonic and biphenyl hydroxyl amin e chloride for a better dispergation. Surprisingly the reactivity of the LPS in the whole blood assay for pyrogen s dropped the more the LPS was dispergated. That's right inverse to the sit uation in the LAL. There are first experiences in the whole blood assay for pyrogens with biol ogical pharmaceuticals. This includes immunoglobulins, vaccines and coagula tion factors. Except of the vaccines and albumin the rabbit assay is the only May to test these pharmaceuticals for pyrogens. The albumins show a little amplification of the reactivity of LPS in the wh ole blood assay for pyrogens. The testing of albumin in the whole blood ass ay for pyrogens showed highly reproproducible results and a detection limit under 50 pg/ml LPS. The immunoglobulins show an amplifying effect as well. Immunoglobulins spik ed with LPS were detected as pyrogenic in all cases. Some vaccines show a slight pyrogenity in the whole blood assay which gets visible when diluting the sample. Undiluted vaccines may cover a LPS contam ination in the whole blood assay for pyrogens.