Comparison of various preparations of nuclear antigens by hemagglutination inhibition (HAI).
A clinical laboratory carrying out tests for antinuclear antibodies requires an efficient, reliable preparation method to produce a high yield of nuclear antigens at low cost and a very sensitive, specific assay method for antigen activity. Various tissues were employed for preparation of small nuclear ribonucleoprotein (snRNP) and Sm antigens for these purposes. Fresh calf thymus cells and nuclei, commercially available calf and rabbit thymus acetone powders, fresh rat kidney and liver cells were used as sources of antigens prepared similarly by methods published previously. Preparations of antigens from whole calf thymus cell extracts were prepared with and without inhibitors to protease and RNase. snRNP and Sm antigens were assayed at each preparation step by hemagglutination inhibition (HAI). Using HAI it was possible to routinely assay snRNP and Sm at nanogram/ml quantities which was 10(6) fold more sensitive than Ouchterlony immunodiffusion. Results were expressed as relative specific activity as compared with calf thymus nuclear extract prepared by conventional methods. Protease and RNase inhibitors did not significantly increase yields. Thymus was the best source of snRNP and Sm. Fresh calf thymus extract produced a good, stable, reliable quantity of antigens, whereas calf and rabbit thymus acetone powders provided antigen at higher specific activity with less labor but slightly lower yields. Thus, considering the total cost of preparations, commercial sources may be superior to fresh sources in the clinical laboratory setting. These studies also revealed the utility of the sensitive HAI test not only in the clinical laboratory but also for further research endeavors.
D000818:Animals; D000974:Antibodies, Antinuclear; D000941:Antigens / Q000302:isolation & purification; D034961:Antigens, Nuclear; D001324:Autoantigens / Q000302:isolation & purification; D002417:Cattle; D006385:Hemagglutination Inhibition Tests; D007668:Kidney / Q000276:immunology; D008099:Liver / Q000276:immunology; D009698:Nucleoproteins / Q000302:isolation & purification*; D011817:Rabbits; D051381:Rats; D012261:Ribonucleoproteins / Q000276:immunology / Q000302:isolation & purification; D017411:Ribonucleoproteins, Small Nuclear; D013950:Thymus Gland / Q000276:immunology; D055517:snRNP Core Proteins
D000974:Antibodies, Antinuclear; D000941:Antigens; D034961:Antigens, Nuclear; D001324:Autoantigens; D009698:Nucleoproteins; D012261:Ribonucleoproteins; D017411:Ribonucleoproteins, Small Nuclear; D055517:snRNP Core Proteins