Preparation and determination of egg white ovomucin
1970
Young, Louis Lee
A simple and rapid method has been developed for producing egg white ovomucin in a pure and natural form. The procedure consists of diluting the egg white sample in an appropriate buffer and applying the diluted sample to a previously equilibrated Sepharose 4B column. Since the molecular weight of ovomucin exceeds the upper exclusion limit of the Sepharose 4B, the ovomucin is eluted with the excluded volume. Ovomucin so produced contained 10.7 mg of sialic acid per 100 mg of protein, was electrophoretically nonmobile on paper electrophoresis and was precipitated by acidification and dilution with distilled water. The preparation was free of lysozyme activity but was precipitated by the addition of active lysozyme. A sample of ovomucin prepared by the gel filtration method and concentrated by ultrafiltration was periodically injected into rabbits to prepare an ovomucin antiserum. The antiserum was used to titrate the ovomucin standards. It was found that as the ovomucin content of the standards decreased, the end point in the titration decreased. The ovomucin concentration in a sample of egg white was then determined by titration against the ovomucin antiserum. An ovomucin concentration of 0.2 percent was observed which is in good agreement with published results.
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