Purification of viruses and viral vectors using OptiPrepTM
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چکیده
1. Comparison with other density gradient media Compared to CsCl and sucrose there are procedural advantages to the use of OptiPrepTM: ♦ OptiPrepTM is a sterile solution of 60% (w/v) iodixanol; it is simply diluted with saline to prepare sterile gradient solutions. It is the only gradient medium manufactured under strict FDA and EU cGMP compliance. ♦ CsCl and sucrose are both toxic to cells. ♦ Iodixanol is non-toxic to cells; it has very low endotoxin levels (<1 EU/ml); measured levels on each batch are usually <0.13 EU/ml. ♦ CsCl must be removed prior to HPLC or gel electrophoresis; iodixanol rarely needs removing prior to further processing, except for electron microscopy studies. ♦ CsCl gradients lead to major reductions in viral infectivity. Virus from iodixanol gradients shows a higher % recovery of infectivity and much lower average particle/infectivity ratios compared to that from CsCl gradients (see Table 1) ♦ Although sucrose is generally less deleterious to viral infectivity than CsCl, it can nevertheless have serious effects on viral structure; in particular the loss of surface glycoproteins from enveloped viruses [1].
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