| Autor: |
Powell, Sharon K., Kaloss, Michele A., Pinkstaff, Anne, McKee, Rebecca, Burimski, Irina, Pensiero, Michael, Otto, Edward, Stemmer, Willem P.C., Soong, Nay-Wei |
| Předmět: |
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| Zdroj: |
Nature Biotechnology; Dec2000, Vol. 18 Issue 12, p1279, 4p |
| Abstrakt: |
Manufacturing of retroviral vectors for gene therapy is complicated by the sensitivity of these viruses to stress forces during purification and concentration. To isolate viruses that are resistant to these manufacturing processes, we performed breeding of six ecotropic murine leukemia virus (MLV) strains by DNA shuffling. The envelope regions were shuffled to generate a recombinant library of 5 × 10[SUP6} replication-competent retroviruses. This library was subjected to the concentration process three consecutive times, with amplification of the surviving viruses after each cycle. Several viral clones with greatly improved stabilities were isolated, with the best clone exhibiting no loss in titer under conditions that reduced the titers of the parental viruses by 30- to 100-fold. The envelopes of these resistant viruses differed in DNA and protein sequence, and all were complex chimeras derived from multiple parents. These studies demonstrate the utility of DNA shuffling in breeding viral strains with improved characteristics for gene therapy. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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