Enhanced purification of platelet beta-lysin



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Texas Tech University


Experimental alterations in the purification scheme established by Donaldson et al. for platelet B-lysin from rabbit serum were made with the objectives of enhanced bactericidal activity and increased stability. Either bovine serum albumin or reduced glutathione, when added to the filtrates in the purification scheme, increased bactericidal activity against Bacillus subtilis and Escherichia coli. In addition, ammonium bicarbonate was a more effective eluant than sodium chloride, which was used in the established procedure for removing 6-lysin from asbestos filter pads. The modified procedure, using glutathione and ammonium bicarbonate, yielded a stable 6-lysin preparation with increased bactericidal activity. Three UV absorbance peaks were obtained after filtration on a Sephadex G-50 column, with the third and largest peak showing substantial bactericidal activity. Bactericidal activity was also demonstrated in g-lysin samples after affinity chromatography on Affi-Gel blue. Analysis of the various 6-lysin preparations by polyacrylamide gel electrophoresis showed that the affinity chromatography step reduced albumin contamination in the samples of platelet 6-lysin.



Electrophoresis, Affinity chromatography, Anti-infective agents, Beta-lysin