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Use of Sodium Polyanetholesulfonate-CaCl₂ for Removal of Serum Nonspecific Inhibitors of Rubella Hemagglutination: Comparison with Other Polyanion-Divalent Cation Combinations

¹ Department of Microbiology and Parasitology, Faculty of Medicine, University of Toronto, Toronto, Ontario, Canada M5S 1A1

ABSTRACT

By using trypsin-treated human type O cells as indicators, we compared the abilities of four polyanion-divalent cation combinations (heparin-MnCl₂; high-and low-molecular-weight dextran sulfate-CaCl₂; and sodium polyanetholesulfonate [SPS]-CaCl₂) for removal of serum non-immunoglobulin (lipoprotein) inhibitors of rubella hemagglutination. The combination of SPS-CaCl₂ was found to be the most effective, precipitating completely the pre-ß and ß-lipoproteins and reducing the -lipoprotein levels by more than 50%. Hemagglutination patterns after this treatment were clear and stable, and, when normal sera were tested, hemagglutination-inhibition (HI) titers were comparable to those obtained after standard heparin-MnCl₂ treatment. High-molecular-weight dextran sulfate-CaCl₂ removed serum lipoproteins almost as effectively as SPS-CaCl₂. However, problems of nonspecific agglutination and the heavy hemagglutination patterns resulting made this combination unacceptable for routine purposes. Neither low-molecular-weight dextran sulfate-CaCl₂ nor heparin-MnCl₂ removed the pre-ß lipoproteins completely, and occasionally traces of ß-lipoprotein also remained after treatment. The presence of pre-ß lipoproteins in normal sera after treatment may be of no consequence in the HI test since we have found that the very-low-density lipoprotein fractions obtained by ultracentrifugal methods from normal sera (those corresponding to the pre-ß fractions obtained by electrophoresis) had no HI activity. However, very-low-density lipoprotein fractions from all hyperlipemic sera tested had HI activity (titers ranging from 1:16 to 1:1,024) which, in the majority of cases, was not eliminated after heparin-MnCl₂ treatment. In every case, treatment with SPS-CaCl₂ removed this nonspecific activity completely. Since hyperlipemic sera may occasionally be encountered in routine rubella HI antibody testing, we recommend the use of SPS-CaCl₂ rather than heparin-MnCl₂ for pretreatment of sera.