Abstract
Mouse L cell interferon induced by Newcastle disease virus was purified by a combination of salt precipitation, ion exchange chromatography, gel filtration, and affinity chromatography using anti-interferon anti-body. The preparation was labeled with 125I at a later step of purification, which served as an index of protein concentration. Two main species of interferon molecules differing in molecular weight were separated from each other, and the final preparations obtained were shown to be essentially pure by polyacrylamide gel electrophoresis in the absence as well as in the presence of sodium dodecyl sulfate. The highest specific activities obtained were 2.6 X 10(9) and 7.3 X 10(8) international units/mg of protein (determined by 125I radioactivity) for the 40,000- and 24,000-dalton species, respectively. The preparations at different steps of purification, including those of the highest purity, were tested for the anti-cell growth activity. Their activities were found to be indistinguishable from each other when compared at the same antiviral doses, indicating that the anti-cell growth activity is carried by the interferon molecules themselves.
Highlights
Two main species of interferon molecules differing in molecular weight were separated from each other, and the final preparations obtained were shown to be essentially pure by polyacrylamide gel electrophoresis in the absence as well as in the presence of sodium dodecyl sulfate
In the present study, which is a continuation of our efforts to purify mouse L cell interferon (1, 15, 16), a number of technical improvements were introduced, including affinity chromatography using anti-interferon antibody of high specificity and radioactive labeling of proteins that greatly increased the sensitivity of protein determinations
Purification of Interferon-The principle of purification was essentially the same as that employed in this laboratory previously (1, 16), except that affinity chromatography was employed and that radioactivities were introduced into the preparation
Summary
Purification of Interferon-The principle of purification was essentially the same as that employed in this laboratory previously (1, 16), except that affinity chromatography was employed and that radioactivities were introduced into the preparation. A detailed description of each step in the procedure is given in the miniprint supplement immediately following this paper. The purity of the F and S interferon preparations obtained was examined by gel electrophoresis in the presence and absence of SDS. Both of the interferons migrated in somewhat broader peaks in these runs than at the earlier steps, and because of some inactivation during the process, the specific activities at the peak fractions were not very high
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