Serum-Agar Double Diffusion Studies with Ascaris Antigens

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Abstract
An antigenic analysis with an agar double diffusion technique was made with Ascaris lumbricoides var suum antigens. Whole worm antigens (saline soluble extracts) were compared to autoclaved antigens and polysaccharide-protein antigens prepared in a variety of methods. Four techniques were assayed for the preparation of polysaccharide antigens. Ethanol precipitation, formamide followed by precipitation with acid alcohol and acetone, boiling in 30% KOH followed by ethanol precipitation, and precipitation with acetone of saline soluble antigens. Polysaccharide-protein antigens were also prepared from Ascaris tissue fractions. Initial boiling of whole worm homogenate with 30% KOH followed by precipitation with ethanol yielded an antigen that showed 1 band in the agar assay. Polysaccharide-protein antigens obtained with ethanol, formamide and acetone precipitation showed 4–9 bands with whole worm homogenate. Tissue polysaccharide-protein antigens were less complex than the whole worm antigen when they were prepared with formamide. The formamide method in general yielded antigens that were immunologically less complex than antigens produced by the other methods (with the exception of the KOH method with the whole worm antigen). Of the 5 bands found in whole worm formamide polysaccharide-protein antigen assayed by the double diffusion method, 1 component was isolated in the cuticle and a second immunological distinct antigen in unembryonated egg. All antigens contained appreciable amounts of protein and carbohydrate and have been tentatively called polysaccharide-protein complexes.

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