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171.
P53 codon 72 polymorphism and longevity: additional data on centenarians from continental Italy and Sardinia
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Bonafè M Olivieri F Mari D Baggio G Mattace R Berardelli M Sansoni P De Benedictis G De Luca M Marchegiani F Cavallone L Cardelli M Giovagnetti S Ferrucci L Amadio L Lisa R Tucci MG Troiano L Pini G Gueresi P Morellini M Sorbi S Passeri G Barbi C Valensin S Monti D Deiana L Pes GM Carru C Franceschi C 《American journal of human genetics》1999,65(6):1782-1785
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Recombinant adenovirus is one of the primary vectors for human gene therapy. However, the aggregation of unstable virus has
been a recurring problem during the production of purified virus for human therapeutics. To facilitate the development of
a robust manufacturing process for recombinant adenovirus vectors, a convenient and reliable size distribution analytical
assay is necessary and we demonstrate here that disc centrifuge sedimentation is applicable to this purpose. Using the disc
centrifuge system and the line start method, the assay can provide particle size distribution of adenovirus samples within
30 min. The assay can detect virus concentrations down to 0.01% (w/v) or 3 × 1011 particles per ml. The apparent hydrodynamic diameter of recombinant adenovirus was determined to be about 0.063 μm. Furthermore,
the disc centrifuge analysis was able to detect adenovirus dimers, trimers, and tetramers, consistent with a rigid sphere
approximation for adenovirus, as well as a large aggregate of 0.35 μm. The appearance of viral aggregates is confirmed by
increased light scattering based on A320/A260 ratios. The technique could be useful for monitoring the kinetics of aggregation for adenovirus and other DNA and RNA viruses
in the submicron region. Therefore, this novel assay provides a critical tool for purification development of viral vectors
for meeting therapeutic and research needs.
Received 18 September 1997/ Accepted in revised form 15 May 1998 相似文献
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B Baggio S Favoro A Antonello A Zen F Zen A Borsatti 《The Italian journal of biochemistry》1975,24(3-4):199-206
The subcellular localization of renin and kallikrein in rat kidney cortex homogenate was investigated using both differential and density gradient centrifugation techniques. Highest specific activity of renin was found in the heavy mitochondrial fraction. Mitochondrial localization of renin was further supported by the behaviour of succinic dehydrogenase. By differential centrifugation, highest specific activity of kallikrein was found in the light mitochondrial fraction, while by density gradient centrifugation kallikrein was almost completely recovered in the lysosomal fraction. Lysosomal localization of kallikrein is further supported by the behaviour of acid phosphatase. The different subcellular localizations of renin and kallikrein are confirmed and the suggestion that kallikrein is located in the lysosomes is advanced. 相似文献
176.
Astorga and Williams1 have demonstrated that leucocytes from some patients with rheumatoid arthritis (RA) have an impaired one-way mixed leucocyte reaction2 (MLR) when stimulated with leucocytes from other such patients. No definite explanation of this phenomenon is given. We have investigated other diseases in which auto-immune aetiology is suspected to see whether similar phenomena are associated with them. Here we report our investigations of multiple sclerosis (MS). 相似文献
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