Purification of Simian Virus 40 and JC T-Antigens From Transformed Cells 2

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Abstract
Simian virus 40 (SV40) T-antigen was purified from an SV40-transformed hamster cell line (H-50) by three successive separation procedures. A crude lysate from transformed cells was chromatographed on a Biogel column under dissociating nondenaturing conditions. A peak of the immunoreactive material was found between the elution volumes of bovine serum albumin and IgG. This material was further purified by ion exchange chromatography on DEAE-cellulose and phosphocellulose columns. After the last purification step, the pooled immunoreactive material derived from SV40-transformed cells had a specific activity increased 166-fold over that of the crude cell lysate. The total recovery was 15.9%. About 15% of the 1251-labeled immunoreactive material from the last purification step could be precipitated with hamster anti-SV40 T-serum. The procedure for purification of SV40 T-antigen described here had distinct advantages over those previously described, but the known lability of the SV40 T-antigen was still an obstacle in the preparation of antigen with greater specific activity. A similar procedure was applied for the purification of JC T-antigen from a JC virus-transformed hamster cell line (HJC-15). JC T-antigen was, however, even more labile than SV40 T-antigen. The elution profiles of the two antigens on the Biogel and DEAE-cellulose columns were similar

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