Use of sodium polyanetholesulfonate-CaCl2 for removal of serum nonspecific inhibitors of rubella hemagglutination: comparison with other polyanion-divalent cation combinations.
By using trypsin-treated human type O cells as indicators, we compared the abilities of four polyanion-divalent cation combinations (heparin-MnCl(2); high-and low-molecular-weight dextran sulfate-CaCl(2); and sodium polyanetholesulfonate [SPS]-CaCl(2)) for removal of serum non-immunoglobulin (lipoprotein) inhibitors of rubella hemagglutination. The combination of SPS-CaCl(2) was found to be the most effective, precipitating completely the pre-beta and beta-lipoproteins and reducing the alpha-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-MnCl(2) treatment. High-molecular-weight dextran sulfate-CaCl(2) removed serum lipoproteins almost as effectively as SPS-CaCl(2). However, problems of nonspecific agglutination and the heavy hemagglutination patterns resulting made this combination unacceptable for routine purposes. Neither low-molecular-weight dextran sulfate-CaCl(2) nor heparin-MnCl(2) removed the pre-beta lipoproteins completely, and occasionally traces of beta-lipoprotein also remained after treatment. The presence of pre-beta 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-beta 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-MnCl(2) treatment. In every case, treatment with SPS-CaCl(2) removed this nonspecific activity completely. Since hyperlipemic sera may occasionally be encountered in routine rubella HI antibody testing, we recommend the use of SPS-CaCl(2) rather than heparin-MnCl(2) for pretreatment of sera.
D000914:Antibodies, Viral / Q000032:analysis*; D000998:Antiviral Agents / Q000302:isolation & purification*; D001557:Benzenesulfonates; D002122:Calcium Chloride; D002712:Chlorides; D002784:Cholesterol / Q000097:blood; D003911:Dextrans; D006385:Hemagglutination Inhibition Tests; D006493:Heparin; D006801:Humans; D006949:Hyperlipidemias / Q000097:blood / Q000276:immunology; D007136:Immunoglobulins / Q000032:analysis; D008074:Lipoproteins / Q000097:blood; D008079:Lipoproteins, VLDL / Q000302:isolation & purification*; D008345:Manganese; D011074:Polyanetholesulfonate; D012409:Rubella / Q000276:immunology*; D014280:Triglycerides / Q000097:blood
D000914:Antibodies, Viral; D000998:Antiviral Agents; D001557:Benzenesulfonates; D002712:Chlorides; D003911:Dextrans; D007136:Immunoglobulins; D008074:Lipoproteins; D008079:Lipoproteins, VLDL; D014280:Triglycerides; D008345:Manganese; D011074:Polyanetholesulfonate; D006493:Heparin; D002784:Cholesterol; D002122:Calcium Chloride