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Article
April 1972

Purification of Large Quantities of Australia Antigen by Density Gradient Centrifugation

Author Affiliations

DVM, Marcy l'Etoile, France
From the Institut Merieux, Service de Recherches, Marcy l'Etoile, France.

Am J Dis Child. 1972;123(4):322-324. doi:10.1001/archpedi.1972.02110100054022
Abstract

In order to prepare specific, animal, anti-Australia immunoglobbulins on a large scale, we have attempted hepatitis-associated antigen (Au-SH) purification from human ascitic fluid, plasma, and sera, with a zonal rotor (MSE B XV), in sucrose gradients.

Accepting the 110S sedimentation coefficient value,1 the estimated centrifugation conditions did not provide a satisfactory separation of Au-SH from largest proteins. Considering this slow rate of zonal sedimentation, we estimated a value at most of 50S. New centrifugation conditions were calculated. The original materials are constituted of 100- to 500-ml batches of positive Au-SH serum undergoing several cycles of purification (18 hours; 25,000 rpm; 5 C on a 15% to 45% w/w sucrose gradient in phosphate Sørensen buffer, pH 7.5). Before each cycle, the Au-SH-positive fractions are filtered on membranes (Amicon PM 30) to provide 100 ml with a 1.03 280 nm 254 nm 10 15 20 EID + 35 Fig 1.—Findings on the

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