PURIFICATION OF A TYPE-5 RECOMBINANT ADENOVIRUS ENCODING HUMAN P53 BYCOLUMN CHROMATOGRAPHY

We have investigated the use of column chromatography for the purifica tion of ACN53, a recombinant adenovirus type 5 encoding the human p53 tumor suppressor protein, Anion exchange, size exclusion, hydrophobic interaction, and metal chelating resins were tested; each was found to have distinct advantages and disadvantages, Based on these data, a ra pid method was devised for the purification of ACN53, The resultant pr oduct was characterized and compared to cesium chloride density-gradie nt purified virus by SDS-PAGE, Western blot analysis, absorbance spect rum, total particle-to-infectious particle ratio, expression of p53 ge ne product in Saos-2 cells, growth inhibition of Saos-2 cells, and con tamination by ATCC-293 host cell proteins, The results show that colum n chromatography offers an alternative to ultracentrifugation for the purification of recombinant adenoviruses for use in human gene therapy trials and other research applications.