Determination of the Optimal Ammonium Sulfate Concentration for the Fractionation of Rabbit, Sheep, Horse, and Goat Antisera

Abstract

Various ammonium sulfate concentrations and reaction conditions were employed in the fractionation of sera from rabbits, sheep, horses, and goats. Precipitates and supernatant fluids were analyzed by electrophoresis to study the effects of the controlled variables. At room temperature, the third precipitate in 35% saturated (NH 4 ) 2 SO 4 was the best fraction from both rabbit and sheep sera; 80 to 90% of the gamma globulins were recovered. The second and third precipitates of horse sera proteins in 30% saturated (NH 4 ) 2 SO 4 were both satisfactory, but only 44% of the gamma globulin was recovered after three precipitations. Goat sera yielded a very satisfactory fraction; 80% of the gamma globulin was recovered after two precipitations—the first in 30% and the second in 45% saturated (NH 4 ) 2 SO 4 . The composition of these fractions was not influenced by the p H of the sulfate solutions ( p H 5.8 and 7.2), by a range of normal room temperatures (20 to 30 C), or by diluting the sera before fractionation. Crude globulins and fluorescein isothiocyanate-labeled globulins were successfully refractionated by one precipitation in the optimal sulfate concentration for the appropriate animal species. The refractionated products contained considerably less beta and alpha globulins than did the original crude fractions and little or no albumin.

Preparation and testing of vaccines prepared from the envelopes of Venezuelan, eastern, and western equine encephalomyelitis viruses

“…Mouse hyperimmune antisera were produced by inoculating animals once every 7 days for 4 weeks intramuscularly with approximately 200 ug of isolated envelope components. Globulin fractions were prepared from these sera using ammonium sulfate precipitation as described by Hebert et al (12).…”

Partial Purification and Characterization of Exo-Polygalacturonase Produced by Penicillium oxalicum AUMC 4153

“…To check for the presence of sulfate, mix a small volume of the dialyzing fluid with an equal volume of a saturated barium chloride solution. If the mixture does not become cloudy, the dialysis can be considered complete [41]. Preparation of dialysis cellulose tubing membrane was carried out as described by Sigma-Aldrich chemical company, Germany.…”

Detection of enteric campylobacteriosis in children

“…Indirect fluorescence determinations were performed with rabbit antiserum and fluorescein-conjugated goat anti-rabbit gamma globulin (10,16,17). Rabbit antisera were prepared against C. fetus subsp.…”